Method of removal of pathologic prion protein from blood products

FIELD: medicine.

SUBSTANCE: blood product filtration through filter, filled with carrier, covered with polymer, is performed. Said polymer is formed by three elements: element derived from hydrophobic polymerised monomer, element, derived from polymerised monomer, which contains main nitrogen-containing part, and element, derived from polymerised monomer, which contains proton neutral hydrophilic part as balance. After filtration collection of blood product is carried out. Efficiency of pathologic prion protein removal constitutes from 1.2 to 4.1 and higher.

EFFECT: efficient removal of pathologic prion protein from blood product with simultaneous removal of leukocytes.

12 cl, 1 dwg, 1 tbl, 23 ex

 

The technical field

The present invention relates to a method for removal of pathological prion protein from a blood product. In particular, the present invention relates to a method of selective removal of pathological prion protein that may be present in the blood product, such as whole blood, a concentrated solution of red blood cells or platelets.

The level of technology

Transmissible spongiform encephalopathy (TSE) or prion disease causes of fatal neurodegenerative diseases in humans and other mammals. The most famous scrapie sheep and spongiform encephalopathy in cattle (BSE). Human diseases include sporadic disease Creutzfeld-Jakob disease (sCJD), iatrogenic disease Creutzfeld-Jakob syndrome of Gerstmann-Straussler-Sheinker (GSS), fatal familial insomnia (FFI) and Kuru. Prion disease can occur as a result of changes in the natural conformation of the normal prion protein in variant pathological prion protein (variant prion protein can cause prion disease), which infects humans and other mammals. A variant of the prion protein has a structure enriched β-layers compared to normal prion protein, so a variant of the prion protein has a high hydrophobicity, easily forms multimeric resistant and p is tease K.

Recently, several works have shown that <variant> CJD, which occurs usually in the UK, associated with consumption of beef cows infected with BSE (non-patent documents 1 and 2). In these works it was assumed that <variant> CJD can be transported by eating beef and transmitted from person to person through blood transfusion <product blood> or migration variant prion protein present in the tissue transplant. In this regard, in 2004, there were described two cases of infection <variant> CJD recipients receiving donor blood, have developed <variant> CJD after giving blood, and in 2006 was described in the third case, so there is a high probability of transfer <variant> CJD by transfusion. Many people infected with CJD, the disease does not develop, and blood products derived from such people can contribute to the spread of infection. Thus, you need a way to remove the pathological prion protein from a blood product.

In the field of blood transfusion is common so-called transfusion of blood containing leukocytes, when transfused blood product after removal of leukocytes contained in the blood. This procedure is carried out, since it was found that side effects such as headache, who osnato, fever and non-haemolytic febrile reaction accompanying a blood transfusion, severe side effects, such as alloantigen sensitization, disease post-transfusion graft-versus-host (GVHD) and viral infection, seriously affecting the recipient, usually due to the presence of leukocytes in the blood product used for transfusion. The filtering method has several advantages such as high efficiency removal of leukocytes, ease of use and low cost, so it is widely used as a method of removing leukocytes from <product blood>. Upon receipt of the product of blood blood is usually filtered in the center of the blood and the last time this procedure is performed before storing for complete quality control of the product of blood that does not contain leukocytes, the filtering method, i.e. using a filter that removes leukocytes. Typically, when the blood is filtered by using a filter that removes leukocytes, blood center, blood bag containing blood product to be filtered, placed on 70-150 cm higher than the package for collecting filtered blood product, and the product of the blood is filtered under gravity.

As systems for the production of blood that does not contain leukocytes, widely used two types of C is the (type SCD and a built-in type). In the case of an application system type SCD package containing the product of the blood to remove leukocytes, aseptically connected to a system for removal of leukocytes. Thus, only connect the filter to remove leukocytes and package for the collection of blood product after filtration. In the case of the integrated system the process of drawing blood from the donor to obtain a product of blood is carried out in an integrated system, so that the blood pack typically contains a solution of the preservative or anticoagulant. For sterilization SCD system is usually applied sterilisation by irradiation at low cost. However, sterilization by irradiation can lead to the decomposition of the solution of the preservative and anticoagulant, so for the embedded system is usually applied sterilization by autoclaving.

The level of hemolysis is one of the indicators of the quality of the product-containing blood erythrocytes. Supply high quality blood product, free of leukocytes containing red blood cells, the level of hemolysis should be less than 0.8% (non-patent document 3).

From the point of view of the efficiency of the blood center, cost and loss of blood product, the method of removing abnormal prion protein from a blood product preferably is a method of simultaneous removal of pathological prion protein and leukocytes.

p> In patent document 1 is disclosed a polymer for coating the filter material for the removal of leukocytes containing the item originating from a hydrophobic polymerized monomer, the element originating from the polymerized monomer containing a basic nitrogen-containing part, and the item originating from a polymerized monomer containing a protonic neutral hydrophilic part as material, high-performance removes leukocytes, but not disclosed and are not prompted removal of the pathological prion protein. Filtering the blood that does not contain leukocytes, before storing in many cases includes both filtration at room temperature, at which the blood is filtered at room temperature for one day after blood sampling, and filtering at a low temperature at which blood is filtered after storage of blood in the refrigerator for 1-3 days. In the case of filtration at room temperature, a greater number of leukocytes seeps through the filter than in the case of chilled blood, despite the short time filtering, while in the case of filtration at low temperature filtration is carried out over a longer time, and perhaps to some extent to prevent the leakage of leukocytes through the filter. However, in patent document 1 has not been studied filtering etc is low temperature.

In patent document 2 is disclosed a method of forming complex prion protein in a biological fluid with a polymer matrix containing a hydrophilic, hydrophobic or amphiphilic functional group, or with a prion-binding substance containing aluminum oxide or silicon oxide. However, this complex is not capable of removing leukocytes, although in the examples of the functional group linked to the resin, and thus in case of actual use in the blood center, you must apply the filter to remove leukocytes. The separate removal of leukocytes and pathological prion protein can increase the loss of blood product, labor costs in the blood center, and cost. In addition, it is known that aluminum oxide and silicon induce activation of the coagulation system, and demonstrate high selective adsorption of proteins, and thus these substances are not suitable for blood product.

In patent document 3 is disclosed a method of removing a prion protein from a random sample of fluid using a device such as a flow-through columns and spherical polymer beads whose surface is coated with an agent forming a complex with prion, such as a metal salt (such as sodium) phospholipases acid. However, the sample shall be exposed to the agent, forming the idea is to, in sufficient time for the complex formation agent that forms a complex with almost all of the pathological prion protein contained in the sample. For example, the sample is incubated at a temperature of approximately 30-45°C (preferably 37°C.) for about 1-16 hours. However, the temperature of 37°C is not suitable for storage of blood product, and in the conventional method of using a filter to remove leukocytes, typically, filtering is used at room temperature or at low temperature. In addition, this method applies the filter on the basis of gravity, so this method is not suitable for removal of pathological prion protein or leukocytes from the blood product.

In patent document 4 is disclosed a method of forming complex prion protein and polymer matrix containing a hydrophilic, hydrophobic or amphiphilic group to remove or detect prion or prion-binding substance, such as aluminum oxide or silicon oxide. However, the prion-binding substance used in the examples are not capable of removing leukocytes, and in the case of separate removal of leukocytes and pathological prion protein exists the problem of increasing loss of blood product, labor costs in the blood center and cost. In addition, it is known that the oxide of al is MINIA and silicon induce activation of the coagulation system, and demonstrate a high selective adsorption of proteins, and thus these substances are not suitable for blood product.

Thus, removal of the pathological prion protein from a blood product required a simple and effective method of removing abnormal prion protein from a blood product, and in addition, it is desirable that such a method provides for simultaneous removal of pathologic prion protein and leukocytes.

[Non-patent document 1] G. Chazot, et al., (1996) Lancet 347: 1181

[Non-patent document 2] R.G. Will, et al., (1996) Lancet 347: 921-25

[Non-patent document 3] Guide to the preparation, use and quality assurance of blood components 9th edition/ Council of Europe Publishing

[Patent document 1] WO 03/011924

[Patent document 2] US 2005/0014196 A

[Patent document 3] JP 2002-539081 A

[Patent document 4] JP 2006-522344 A

Description of the invention

Objectives of the invention

Due to the above problems of the conventional technologies, the present invention is to develop a simple and effective way to remove pathological prion protein from a blood product and method for the simultaneous removal of pathological prion protein and leukocytes.

Solutions to problems

The authors of the present invention have conducted extensive studies, which found that pathological prion protein is probably very simple and effective is actively removed from the product of blood through the filter, covered terpolymers, including hydrophobic polymerized monomer, polymerized monomer containing a basic nitrogen-containing part, and the polymerized monomer containing a protonic neutral hydrophilic part (naprimer, including any one of these monomers), and thus completed the present invention. Thus, the present invention relates to:

(1) the method of removal of the pathological prion protein from a blood product, comprising: filtering the blood product through a filter filled with a carrier coated with the polymer formed by three elements, including a 20 molar% or more and 40 molar percent or less of the item originating from a hydrophobic polymerized monomer, 5 molar% or more and 13 molar percent or less of the element originating from the polymerized monomer containing a basic nitrogen-containing part, and the element originating from the polymerized monomer containing a protonic neutral hydrophilic part as of the balance sheet; and collecting the filtered blood product;

(2) a method of removing abnormal prion protein from a blood product according to the above item (1), wherein the blood product is a product of whole blood, and the filter is subjected to sterilization by irradiation and then sterilized and what carlaviruses;

(3) a method of removing abnormal prion protein from a blood product according to the above item (2), characterized in that irradiation is a γ-rays or electron beams;

(4) the removal of abnormal prion protein from a blood product in any of the above items (1)to(3), characterized in that the polymer is a vinyl polymer type;

(5) the method of removal of the pathological prion protein from a blood product in any of the above items (1)to(4), characterized in that the hydrophobic polymerized monomer, polymerized monomer containing a basic nitrogen-containing part, and the polymerized monomer containing a protonic neutral hydrophilic part, are derivatives of acrylic acid and/or derivatives of methacrylic acid;

(6) the method of removal of the pathological prion protein from a blood product in any of the above items (1)to(5), wherein the basic nitrogen-containing group is a tertiary amino group;

(7) the method of removal of the pathological prion protein from a blood product in any of the above items (1)to(6), characterized in that the proton neutral hydrophilic group is a hydroxyl group;

(8) the method of removal of the pathological prion protein from the product kr is VI in any of the above items (1)to(7), characterized in that the filter is a filter for removing leukocytes;

(9) the method of removal of the pathological prion protein from a blood product according to the above item (8), characterized in that the carrier coated with the polymer, is a fibrous material or a material with a spongy structure;

(10) the method of removal of the pathological prion protein from a blood product according to the above item (8) or (9), characterized in that the specific area of the carrier coated with the polymer is 1.0 m2/g or more and 5.0 m2/g or less;

(11) the method of removal of the pathological prion protein from a blood product in any of the above items (8)to(10), characterized in that the average pore diameter of the carrier is 1 μm or more and 60 μm or less;

(12) the method of removal of the pathological prion protein from a blood product in any of the above items (8)to(11), characterized in that the density of the carrier coated with the polymer, 0.1 g/cm3or more and 0.5 g/cm3or less;

(13) the method of removal of the pathological prion protein from a blood product in any of the above items (8)to(12), characterized in that the porosity of the carrier coated with the polymer is 60% or more and 90% or less;

(14) the method of removal of the pathological prion protein isproduct blood in any of the above items (8)to(13), characterized in that the carrier coated with the polymer is a non-woven material; and

(15) the method of removal of the pathological prion protein from a blood product according to the above item (14), characterized in that the fiber diameter of the nonwoven material is 0.3 μm or more and 3.0 μm or less.

The effect of the invention

According to the present invention, it is possible to simply and effectively remove the pathological prion protein from a blood product and simultaneously to remove the pathological prion protein and leukocytes. In addition, the method according to the present invention can be used to obtain high-quality blood product, which has high fluidity and containing less hemolyzed cells.

Brief description of drawings

The drawing shows a schematic representation of a system by which the present invention is.

Description of positions on the drawing:

1: the package with the product of the blood,

2: hose,

3: filter

4: the package for collection of blood product after filtering.

The best option of carrying out the invention

Below the present invention is described in more detail.

The term "polymer" in the present invention means a polymer (polymers), including the item originating from a hydrophobic polymerized monomer, the element originating from Polimeri what has been created monomer, containing basic nitrogen-containing part, and the item originating from a polymerized monomer containing a protonic neutral hydrophilic part.

The term "element" in the present invention means the minimum recurring item originating from each of the polymerized monomer in the composition of the polymer molecules. For example, in the case of polyaddition of the polymerized monomer of the vinyl compound with the formula CH2=CXY (X: H or Deputy, other than N, Y: Deputy, different from X) by disclosing the double bond of the minimum repeating element represents -(CH2-CXY)-. When the polymer is synthesized by polycondensation of the polymer precursor with the formula A-(R)-B (R: part of, not released during polymerization, a and b: part released during the polymerization reaction), as the minimum repeating element may be made -(R)-when a and b are released during polymerization.

Examples of hydrophobic polymerized monomer, particularly from the point of view of accessibility and ease of use include: steren; methylstyrene; acrylates and methacrylates such as methyl acrylate, methyl methacrylate, acrylate, methacrylate, butyl acrylate, butylmethacrylate, phenylacrylate, fenilsalicilat, hexyl acrylate, ethylhexylacrylate, trichlorethylene trichlorethylene; alkenes, such as pentan, hexene, hapten and octene; an organic silicon compound such as silicone and siloxane; and organofluorine curable monomers, in which at least one hydrogen atom in the composition of the ethylene is replaced by a fluorine atom. However, hydrophobic polymerized monomer is not limited to the substances listed above. Of these monomers, from the viewpoint of availability and ease of use of the preferred monomers containing a vinyl group in the quality of the polymerized by the polymerization (vinyl polymerization) is formed of a polymer of the vinyl type. Further preferred hydrophobic polymerized monomers are derivatives of acrylic acid and derivatives of methacrylic acid. The most preferred hydrophobic polymerized monomers are acrylates and methacrylates.

Materials containing basic nitrogen-containing functional group, have a positive charge on the surface in a biological fluid, which has the effect of increasing the efficiency of removal of pathological prion protein. The term "polymerized monomer containing a basic nitrogen-containing part"in this invention means the polymerized monomer containing a basic nitrogen-containing part, described below. As the basic nitrogen-containing groups may you is to tapati primary amino group, secondary amino group, tertiary amino group, Quaternary amino group, peridroma group, imidazolone group and others. Preferred basic nitrogen-containing amino group is a tertiary amino group. The polymerized monomers containing basic nitrogen-containing part from the point of view of availability and ease of use, include vinyl derivatives of nitrogen-containing aromatic compounds, such as vanillin, 2-vinylpyridine, 4-vinylpyridine, 2-methyl-5-vinylpyridine, 4-vinylimidazole, N-vinyl-2-ethylimidazole, and N-vinyl-2-Mei; acrylates and methacrylates, such as dimethylaminoethylacrylate, dimethylaminoethylmethacrylate, diethylaminoethylmethacrylate, diethylaminoethylmethacrylate, 3 - dimethylamino-2-hydroxypropylamino and 3-dimethylamino-2-hydroxypropylmethacrylate; amide derivatives of acrylic acid and methacrylic acid, such as amide and N,N - dimethylaminoethylacrylate acid amide and N,N-dimethylaminoethylmethacrylate acid amide and N,N - diethylaminoethylcellulose acid amide and N,N - diethylaminoethylmethacrylate acid and amide N,N-dimethylaminopropylamine acid; derivatives of styrene, such as p-dimethylaminomethylene and p-diethylaminoacetate; and derivatives, such as Quaternary ammonium salts obtained by reaction of the polymerized monomer with alkylhalides group. One is about the polymerized monomer, containing basic nitrogen-containing part, is not limited to these substances. Of these monomers, from the viewpoint of availability and ease of use, preferred monomers containing a vinyl group as the polymerized part forming the vinyl polymer type by polymerization (vinyl polymerization). As a polymerized monomer containing a basic nitrogen-containing part, the preferred derivatives of acrylic acid and methacrylic acid. More preferred as the polymerized monomer containing a basic nitrogen-containing part, acrylates and methacrylates. Of these substances are particularly preferred dimethylaminoethylacrylate, dimethylaminoethylmethacrylate, diethylaminoethylmethacrylate and diethylaminoethylmethacrylate.

The term "polymerized monomer containing a protonic neutral hydrophilic part"in this invention means a monomer whose depolimerizatia part dissociates, releasing protons (H+), and a monomer not having a strong acidity or basicity as carboxylic acid or the basic amino group. The polymerized monomer containing a protonic neutral hydrophilic part, shows large hydrophilic properties compared with the monomer containing nephratonia neutral hydrophilic part, and has a lovely view the ACLs abilities to activate and prevent product flow of blood. Examples of the protonic neutral hydrophilic part include a hydroxyl group, aldehyde group with the proton in α-position and an amide group having a proton in α-position and 1,3-dicarbonyl group. As depolymerizing protonic neutral hydrophilic part is particularly preferred hydroxyl group. Examples of the polymerized monomers containing neutral hydrophilic part, include 2-hydroxyethylmethacrylate, hydroxypropylmethacrylate, hydroxyethylacrylate, acrylamide and methacrylamide. However, the polymerized monomer containing a protonic neutral hydrophilic part, are not limited to the above substances. Of these monomers, from the viewpoint of availability and ease of use, preferred monomers containing a vinyl group as the polymerized part forming the vinyl polymer type by polymerization (vinyl polymerization). Of these monomers as polymerized monomer containing a protonic neutral hydrophilic part, the preferred derivatives of acrylic acid and derivatives of methacrylic acid. As a polymerized monomer containing a protonic neutral hydrophilic part, the most preferred acrylates and methacrylates. The term "vinyl polymer-type" in the present invention means poly is EP vinyl type in a broad sense, whose main chain contains no cycles. Specific examples include α-substituted acrylic acid and its derivatives, polyvinyl ether, polyvinyl alcohol, polyvinyl ester, polystyrene and its derivatives and copolymers comprising these polymers, as described in J. Brandrup; E.H. Immergut. 1989 “Polymer Hand book Third Edition, A Wiley-interscience publication, p. VII-5 VII-18.

Removal of pathological prion protein from a blood product containing the component plasma, it is necessary to use the above-mentioned polymer, comprising three elements, including the element originating from a hydrophobic polymerized monomer, the element originating from the polymerized monomer containing a basic nitrogen-containing part, and the item originating from a polymerized monomer containing a protonic neutral hydrophilic part. Even when using a polymer containing only one of these elements, it is impossible to remove the pathological prion protein with high efficiency, and in particular it is impossible to effectively remove the pathological prion protein from a blood product containing the component plasma.

Pathological prion protein contains three different binding region binding to the positively charged functional groups, negatively charged functional groups and hydrophobic function the national groups. It was shown that the isoelectric point of the pathological prion protein corresponds to a value of pH of 4.6, and the pH of the blood product is within approximately 5-7,5, so that the pathological prion protein in the blood product is charged negatively. Thus, the monomer containing a basic nitrogen-containing part and a hydrophobic polymerized monomer, can remove the abnormal prion protein from a blood product with high efficiency. In addition, it is known that the contact between the material bearing negative charges, and the product of the blood can induce the production of bradykinin, causing anaphylactic reaction, such as a fall in blood pressure, a surge of blood to the face, conjunctival hyperemia, smooth muscle contraction or pain, thus, the polymer bearing negative charges, is not suitable for coating the surface of the carrier when used for filtration of the blood product.

On the other hand, the filtering of the blood product using a filter necessary for receiving the blood product of the same quality as a product of blood, obtained by filtering through a conventional filter to remove leukocytes. Protonic neutral hydrophilic part of the polymer necessary to ensure wettability required to distribute product blood through the filter, in which osobennosti to conduct uniform activation of the column, that is, the procedure of filling of the filter product of blood in the first stage of filtration. If the composition of the monomer containing a basic nitrogen-containing part and a hydrophobic polymerized monomer in the polymer exceeds certain levels, it causes a loss of quality, leading to hemolysis or increased filtration time. Thus, for the production of blood, from which the pathological prion protein is removed with high efficiency, it is necessary to adjust the composition of the hydrophobic polymerized monomer polymerized monomer containing a basic nitrogen-containing part, and the polymerized monomer containing a protonic neutral hydrophilic part, in the appropriate limits.

The term "hydrophobic polymerized monomer" in the present invention means the polymerized monomer, has a very low affinity to water and the solubility in water 12 g per 100 g of water or less at 20°C and means a monomer whose molecule contains no basic nitrogen-containing part or proton neutral hydrophilic part. If the solubility is more than 12 g per 100 g of water, can not be achieved high removal efficiency of pathogenic prion protein in the present invention. Preferred solubility in the range of 2 g per 100 g of water or less. The solubility of m which can be determined in a known manner, such as the method of determining the dew point, thermal analysis, electrical method, including the measurement of electromotive force or electrical conductivity of the solution, gas chromatography or indicator method, when the polymerized monomer is a solid. When the polymerized monomer is a liquid, the solubility can be determined using the same techniques that are applicable to solid polymerized with the monomer, as well as the methods known from the prior art, such as capacitive method, the method of light scattering or the way the saturated vapor pressure. As a more simple way, if the boiling point of the polymerized monomer is considerably above the boiling point of water, can be used the way, where from a saturated aqueous solution of the polymerized monomer is evaporated the water and measure the mass of the dry residue.

To achieve greater efficiency in the removal of abnormal prion protein and removal of leukocytes preferably, the polymer included the above monomers in the following concentrations (molar fraction): hydrophobic polymerized monomer, 20 molar% or more and 40 molar percent or less, of the polymerized monomer containing a basic nitrogen-containing part, 5 molar% or more and 13 molar percent or is the Eney; the polymerized monomer containing a protonic neutral hydrophilic part, the concentration resulting from subtracting the sum of the mole fractions of hydrophobic polymerized monomer in the polymer and the polymerized monomer containing a basic nitrogen-containing part, from 100 molar percent. If the molar fraction of hydrophobic polymerized monomer in the polymer is less than 20 molar percent or concentration of the polymerized monomer containing a basic nitrogen-containing part, less than 5 molar%, the removal efficiency of pathological prion protein may not be increased, which is undesirable. If the concentration of hydrophobic polymerized monomer in the polymer exceeds 40 molar percent, may decrease the wettability to the product of the blood stored in the refrigerator, which is undesirable because it causes a decrease in the rate of filtration of the blood product during filtration of blood product using a filter coated with the polymer of the present invention. If the concentration of the polymerized monomer containing a basic nitrogen-containing part, exceeds 13 molar percent, the product of the blood stored in the refrigerator may cause hemolysis, which may be undesirable. In order to achieve even greater efficiency of removal of pathological prion protein and removal of leucocytes Ave is doctitle, to the polymer included the above monomers in the following concentrations: hydrophobic polymerized monomer, 25 molar percent or more and 35 molar percent or less, of the polymerized monomer containing a basic nitrogen-containing part, 7 molar percent or more and 12 molar percent or less, of the polymerized monomer containing a protonic neutral hydrophilic part, in concentrations resulting from subtracting the sum of the mole fractions of hydrophobic polymerized monomer in the polymer and the polymerized monomer containing a basic nitrogen-containing part, from 100 molar percent. Most preferably, the polymer included hydrophobic polymerized monomer at a concentration of 27 molar% or more and 33 molar percent or less, of the polymerized monomer containing a basic nitrogen-containing part, the composition of copolymerization 8 molar percent or more and 11 molar percent or less, of the polymerized monomer containing a protonic neutral hydrophilic part, as of the balance sheet.

The composition of the monomers in the polymer composition can be determined in accordance with generally accepted physicochemical technique. Examples of physico-chemical techniques to determine the composition of copolymerization include known methods such as nuclear magnetic resonance JV is ctroscopy (NMR, -The 1H-13C) and the method of pyrolysis GC/MS. It is also possible to determine whether passes polymerization in accordance with the plan in respect of the composition of the charged monomer or are variations from batch to batch. It is also possible to dissolve and extract the polymer, covering the media, using a solvent for the polymer, and it is possible to analyze the composition of the monomers from the polymer using the above methods. It is also possible to apply the method of the dissolution medium and the polymer present on the surface, deuterated solvent and determine the composition using nuclear magnetic resonance spectroscopy (NMR, -1H, -13C) as a way of determining the composition of the copolymerization. The molecular weight of the polymer can be measured using a known method, gel permeation chromatography. The average molecular weight is preferably in the range of 50,000 or more and 3000000 or less, more preferably 100000 or more and 2000000 or less, most preferably 150000 or more and more than 1,500,000 or less. If the average molecular weight (Mw) less than 50,000, may be undesirable elution of polymer in the product of blood in the uninstall process of pathological prion protein and the removal of leukocytes from the blood product. If the molecular weight (Mw) greater than 3000000, there is an undesirable decrease in rastvorimo and polymer in the solvent, used for coating. In addition, it may be possible stable production of the polymer during polymerization. The polymer may be a random copolymer or a block copolymer. However, the preferred random copolymer, as the block copolymer tends to decrease the solubility of the solvent in the coating and may have a tendency to deteriorate the uniformity of the coating due to the formation of micelles in solution. The polymer can be a linear polymer or a branched polymer. However, the preferred linear polymer chain, since the branched polymer chain may tend to decrease the solubility of the solvent in the coating and may have a tendency to deteriorate the uniformity of the coating due to the formation of micelles in solution. For the synthesis of the polymer may be used the conventional method of polymerization. Can be applied additive polymerization (vinyl polymerization), which is a chain reaction; isomerization polymerization; and the elimination reaction, polymerization, polycondensation, additive polycondensation and similar ways, which is a sequential reaction. As the chain transfer upon receipt of the polymer can be used radicals or ions. As for the type of polymerization, as an exampl which can lead to polymerization in solution, the polymerization mass deposition with simultaneous polymerization, emulsion polymerization. Of these methods, preferred polymerization in solution.

The following is an example of the method of polymerization.

As a solvent for polymerization using ethanol, and ethanol, which was dissolved monomers and initiator (diazo-compound), is added dropwise to the reaction, and the solution is then stirred at a constant temperature equal to or lower than the boiling point of ethanol, under nitrogen atmosphere. If necessary, may be added stabilizer or other substances. The yield of the reaction is measured and confirmed by using a conventional method such as gas chromatography. The possibility of purification of the reaction mixture using conventional method of chemical cleaning to remove impurities such as low molecular weight components and unreacted substances contained in the polymer or in the reaction solution containing the polymer, which can be eluted during processing of the product. As the purification method, you can lead the way in which the reaction mixture is poured into a solvent, solvent impurities, but does not dissolve the polymer, and then separating the precipitate by filtration or decantation. Alternatively, you can lead the way, where the precipitate is washed with a solution of the Telem solubility slightly bigger than the solubility of the solvent containing the precipitate (for example, a mixture of solvent and solvent-containing sludge), and dry the residue under reduced pressure until such time as the sludge does not become constant.

There is no specific limitation to the type of medium, provided that the material has pores through which can be filtered blood. Among the different conformations of carriers suitable for use, particularly preferred fibrous materials such as natural fiber, glass fiber, knitted, woven or non-woven fabric, porous membrane, and the material having a spongy structure that contains the extended three-dimensional network of pores. Various media, such as organic polymeric materials, inorganic polymeric materials and metals, can be used without restriction provided that blood cells are not damaged. Of these materials, the preferred organic polymeric materials due to its excellent technological properties such as cutting. For example, polyester, polyolefin, polyacrylonitrile, polyamide, polystyrene, polymethylmethacrylate, polivinilhlorid, polyurethane, polyvinyl alcohol, polyvinyl acetate, polysulfone, polyvinylidene fluoride, politicalparties, a copolymer of vinylidenefluoride and tetrafluoroethylene, polyesterol is h, polyacrylate, a copolymer of butadiene and Acrylonitrile, a block copolymer of polyester and palamida, cellulose and cellulose acetate. However, the carrier of the present invention is not limited to the above examples. Preferred polyester and polyolefin, and particularly preferred polyester.

The term "carrier coated with the polymer" in the present invention means a medium obtained by fixing the polymer to the surface of the carrier so that the polymer is not aluinum in the product during processing of the product. As the method of fixation of the polymer on the surface of the carrier can be applied chemical method of forming covalent bonds or physical-chemical method of forming non-covalent bonds. Preferably the amount of polymer 0.6 mg/m2or more and 83 mg/m2or less. When the amount of the polymer is less than 0.6 mg/m2per unit area on the entire surface area of the carrier is undesirable decrease in the efficiency of removal of pathological prion protein and removal of leukocytes; when the amount of polymer in excess of 83 mg/m2that may be undesirable fluctuations filter performance due to uneven coating. The preferred amount of polymer is 5 mg/m2or more and 50 mg/m2or less per unit area on all the surface area of the carrier, particularly preferred amount of the carrier is 10 mg/m2or more and 40 mg/m2or less. The amount of polymer on the surface of the carrier may be determined according to the conventional physico-chemical techniques. As a measure of excess polymer on the surface of the media, you can lead the way in which the coated carrier and the polymer present on the surface, dissolved in deuterated solvent and determine the number using the method of nuclear magnetic resonance (NMR, -1H, -13C). In the description of the present invention in some cases, the coated carrier" refers to a carrier coated with the polymer.

As a method of coating a polymer carrier according to the present invention can lead to known methods, such as fixing the above-mentioned polymerized monomers of the polymer on the carrier by chemical covalent binding (e.g., vaccination), the method of fixation by non-covalent physical-chemical bonding (ionic bond forces van der Waals forces etc) (e.g., floor) and the method of embedding polymer. A more accurate way of direct inoculation of the polymerized monomers or polymer on the surface of the carrier by way of grafting polymerization, such as radiation grafting or plasma grafting or a method of coating a polymer surface esitalapram saturation of the media with a solution of the polymer or causing the platen and the transfer of the conductor on the surface of the carrier are preferred due to the relatively simple production process, as a result of which consistently turn out products with excellent performance. For coating polymer of the present invention the carrier may be used various ways without particular restrictions, provided that the surface of the carrier can be covered fairly evenly, and the pores of the media is not clogged. Examples of methods of coating a polymer carrier include, but are not limited to the method of saturation of the media with a polymer solution, a method of spraying the polymer solution onto the carrier, and method of application or transfer of the polymer solution on a carrier using a roller to fotogravirovka. Of these methods, the preferred method of saturation of the media with a solution of polymer and compression of the medium and the method of application or transfer of the polymer solution on a carrier using a roller to fotogravirovka thanks to the wonderful possibilities of continuous production and low cost. There is no special limitation on the types of solvents that can be used for dissolving the polymer, provided that the solvents do not dissolve the media. Examples thereof include solutions containing water and inorganic salts; alcohols such as methanol, ethanol, propanol and butanol; ketones, such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate; hydrocarbons such the AK benzene and cyclohexane; halogenated hydrocarbons such as chloroform and dichloromethane; sulfur-containing solvents such as dimethyl sulfoxide; amides such as dimethylformamide and dimethylacetamide; and mixtures of different kinds of the above solvents within the limits of their solubility. However, the solvents applicable as solvents, dissolving the polymer of the present invention is not limited to a high degree the above examples. For drying the polymer solution after coating can be used a method which involves the removal of excess solvent by mechanical compression or injection of gas, such as air or nitrogen, and the abandonment of the coated carrier in dry air or under reduced pressure at atmospheric temperature or when heated. To increase the adhesion of the polymer according to the present invention to the carrier surface of the carrier may be treated prior to coating with a suitable agent such as acid or alkali, or irradiated plasma. Adhesion of the polymer to the carrier can be further improved by heat treatment after coating the polymer or by using post-processing, which consists in irradiation of the coated surface rays such as γ-rays or electron rays. Stage coating can be carried out in the production process of the carrier or after manufacture of wearing what I am.

It is known that the physical structure of the conductor plays an important role in the removal of abnormal prion protein and leukocytes. To increase the efficiency of removal of pathological prion protein and leukocytes important factor is the choice of media. With regard to the physical structure of the coated carrier, the specific surface area is 1.0 m2/g or more and 5.0 m2/g or less, preferably 1.1 m2/g or more and 3.0 m2/g or less, and most preferably 1.3 m2/g or more and 2.0 m2/g or less. If the specific surface area of the carrier is less than 1.0 m2/g, it is difficult to achieve high efficiency of removal of leukocytes. If the specific surface area exceeds 5.0 m2/g, it becomes impossible to stable production covered by the media. In practice, when processing a product using a filter for the blood it is preferable that two or more coated media with different values of specific surface area were located so that the specific surface area of the carrier is increased toward the output port.

In relation to the physical structure of the coated carrier preferably, the porosity was 65% or more and 90% or less, more preferably 75% or more, 88% or less. The por is the porosity less than 65% decreases the speed of filtration of blood, and removal of pathological prion protein and leukocytes requires a greater amount of time. At the same time, if the porosity exceeds 90%, the number of intersection points of the fibers, which can be linked pathological prion protein and leukocytes, small, resulting in low efficiency of removal of pathological prion protein and removal of leukocytes. When using a fibrous material, such as non-woven material, as covered by the media, the average fiber diameter is 0.3 μm or more and 3.0 μm or less, preferably 0.5 µm or more and 2.5 μm or less, more preferably 1 μm or more and 2 μm or less. In practice, when processing a blood product using a filter for the blood it is preferable that two or more coated media with different values of the average diameter of the fibers were arranged so that the average diameter of the fibers of the medium decreased toward the output port. In practice, when processing a blood product using a filter for the blood, from the input port from the covered carrier may not necessarily be the media with average fiber diameter of 10 μm or more and 40 μm or less, in order to remove small units.

The average pore diameter mean value (the average diameter of pores, MFP), obtained by analysis is and sample mass of about 50 mg using Porometer Coulter R production Coulter Electronics. The average pore diameter is 1 μm or more and 60 μm or less, preferably 1 μm or more and 30 μm or less, more preferably 1 μm or more and 20 μm or less. When the average pore diameter less than 1 μm is not desirable, the product of whole blood is difficult to penetrate through the pores, at the same time, if the average pore diameter of greater than 60 μm, it may desirable to decrease removal efficiency of leukocytes. In practice, when processing a blood product using a filter for the blood, it is preferable that two or more coated media with different values of the average diameter of the pores were arranged so that the average diameter of pores of the carrier is decreased toward the output port. In practice, when processing a blood product using a filter for the blood from the input port from the covered carrier may not necessarily be the media with an average pore diameter of 50 μm or more and 200 μm or less, in order to remove small units.

In practice, when processing a blood product using a filter for the blood from the side of the output port from the covered carrier may not necessarily be the media with an average pore diameter of 50 μm or more and 200 μm or less, for the purpose of preventing unevenness of the stream.

When filling the container of fibrous material for removal the Oia pathological prion protein and leukocytes density is preferably 0.1 g/cm 3or more and 0.5 g/cm3or less, more preferably 0.1 g/cm3or more and 0.3 g/cm3or less. Method of measurement of density of filling described through example. Non-woven material designed to fill, cut into pieces and measure the padding (cm3) and mass (g). The density can be calculated from the length (cm) material in a real container.

If the average fiber diameter less than 0.3 μm or the average pore diameter less than 1 μm or a density less than 0.5 g/cm3the filter can become clogged with blood cells, or may increase the pressure loss. If the average fiber diameter of more than 3.0 mm, the average pore diameter greater than 60 microns or density less than 0.1 g/cm3may it is undesirable to decrease the removal efficiency of pathological prion protein and removal of leukocytes.

A porous membrane or a material having a foam structure containing three-dimensional network long been used as a coated carrier preferably has an average pore diameter of 1 μm or more and 60 μm or less, preferably 5 μm or more and 50 μm or less, more preferably 10 μm or more and 40 μm or less. If the average pore diameter of less than 1 μm, the filter can become clogged with blood cells or may not be desirable to called loss Yes the population. If the average pore diameter greater than 60 μm, it is undesirable decreases the removal efficiency of leukocytes.

The term "pathological prion protein" in the present invention means a variant of the prion protein, which is known that it causes a disease called spongiform encephalopathy, in humans and animals. Pathological prion protein has the same amino acid sequence as the normal prion protein, but has an altered conformation, enriched β-layers, and resistant to protease K. Pathological prion protein can infect animals, which leads to the development of scrapie, which is an infectious degenerative disease of the nervous system of sheep and goats, and BSE in cattle. In the case of human pathological prion proteins can cause spontaneous disease Creutzfeld-Jakob disease (sCJD), iatrogenic disease Creutzfeld-Jakob syndrome of Gerstmann-Straussler-Sheinker (GSS), fatal familial insomnia (FFI), Kuru and variant CJD, which is transmitted through meat infected with BSE. Pathological prion proteins used in the present invention include all types of pathological prion protein, which can cause any or all of the above disease in any animal, especially humans and domestic animals. In fact, according to the NSS report Samuel Coker at Cambridge Healthtech Institut's 10 thAnnual Transmissible Spongiform Encephalopathies - The Definitive American TSE Meeting in March 2006, the pathological prion proteins derived from a variant of scrapie and spontaneous CJD CJD, can be removed using a filter (LAPRF, production Pall) equally, there was no significant difference in the coefficients of removal. In line with this, we can assume that the protein patterns of pathological prion protein in terms of removal efficiency using the filter is almost the same even in different species of animals or in various diseases, and that the ratio of removal of the pathological prion protein derived from human material, can be calculated based on the ratio of removal of the pathological prion protein derived from animal material.

The term "filter" in the present invention means a product obtained by filling the container for blood product having input and output ports, covered by the media according to the present invention, can also be a medium, catching small units, input or output side of the blood product.

The container material may be a solid resin or a flexible resin. In the case of solid resin examples may include phenolic resin, acrylic resin, epoxy resin, formaldehyde resin, urea resin, Kremneva the resin (silicone), Acrylonitrile-butadiene-stearinovuyu resin, nylon, solid polyurethane, polycarbonate, vinyl chloride, polyethylene, polypropylene and polyester, etc. In the case of a flexible resin may be used a material obtained by forming inlet and outlet port for the product of blood on the sheets of flexible synthetic resin, with subsequent bonding of the resulting parts on the periphery, or molded cylindrical product having input and output ports for blood product. When the material is attached to the container and covering the carrier, the preferred materials having similar thermal and electrical properties. Examples of suitable materials include polyolefins, such as soft polyvinyl chloride, polyurethane, a copolymer of ethylene and vinyl acetate, polyethylene and polypropylene, thermoplastic elastomers such as hydrogenated copolymer of styren butadiene styren, the copolymer steren-isoprene-stiren or its hydrogenated products, and a mixture of a thermoplastic elastomer and a plasticizer, such as a polyolefin or an ethylene-acrylate or the like. These materials are preferred soft polyvinyl chloride, polyurethane, a copolymer of ethylene and vinyl acetate, polyolefin and a thermoplastic elastomer containing them as a main component, particularly preferred soft polyvinyl the reed and the polyolefin.

There is no special restrictions on the form of the container, provided that the container has a shape that includes an input port for a product of blood and an output port for blood product, which were removed pathological prion protein and leukocytes, but preferably form depends on the form of media. For example, if the use of media in the form of a plate, the container may have a flat shape, including polygonal shape such as the shape of a quadrangle or a hexagon, or a rounded shape, such as round shape or oval shape. More precisely, the container includes an input container, comprising an input port for blood product and output container comprising an output port for blood product, and has the form, where both of the container adjacent to the media on both sides directly or through the support body and divide the inside of the carrier into two parts, thus forming a flat filter. Alternatively, when using the media of cylindrical shape is preferred cylindrical shape of the container. More precisely, the container includes a cylindrical body, in which the packaged media, the input end of which includes an input port for a product of blood, and the output end includes an outlet port for blood product. The container has the shape, the inside of which is divided into two parts by about the ECCA fill so that fluid introduced through the inlet port, flows from the outer periphery to the inner periphery (or from the inner periphery to the outer periphery) of the cylindrical carrier, thus forming a cylindrical filter.

The term "blood product" in the present invention is generally defined as a fluid, including whole blood, which may contain pathological prion protein, or one or more blood components derived from whole blood, or a liquid obtained by adding to the liquid anticoagulant or preservative solution. Specific examples include but are not limited to the product of a whole blood product red blood cells, platelets, plasma product, washed suspension of red cells, thawed concentrated solution of red blood cells, synthetic blood, enriched in plasma and erythrocytes leucocytes film.

The term "product whole blood" in the present invention means a product of blood, obtained by adding a solution of the preservative or anticoagulant, such as citrate-phosphate-dextrose (CPD), citrate-phosphate-dextrose-adenine-1 (CPDA-1), citrate-phosphate-2-dextrose (CP2D), citrate dextrose formula A (ACD-A), citrate dextrose formula B (ACD-B) or heparin, to whole blood obtained from a donor.

Below is described a method of removing abnormal prion b is the left main coronary artery from a blood product according to the present invention, paying particular attention to procedures. First described embodiment of the method of obtaining various blood products, which intentionally does not limit the present invention.

(The product of whole blood that does not contain pathological prion protein)

To the collected whole blood add the preservative solution of anticoagulant, such as CPD, CPDA-1, CP2D, ACD-A, ACD-B or heparin and the resulting solution is filtered using a filter according to the present invention, removal of the pathological prion protein from whole blood, thereby obtaining the product of whole blood that does not contain pathological prion protein.

Product whole blood is filtered through a system that includes at least a package containing the product of the blood, the filter pack for the collection of blood product that does not contain pathological prion protein, is aseptically connected to hoses in the specified order (see drawing). The system can be connected to the needle for blood collection, the package for blood collection, the package for the separation of components after centrifugation or hose to intake blood product remaining in the filter. Pathological prion protein can be removed by filtering carried out by the supply of blood to filter through the hose from the package containing the product of blood, placed higher than the filter, using the force of gravity is the input, or by wire product of blood through the device, such as a pump, from the input port of the filter under high pressure and/or from the output port of the filter under reduced pressure. The filtering method can be applied not only to products of whole blood, but also to other blood products.

In the case of removal of the pathological prion protein to the store to obtain product of blood that does not contain pathological prion protein, pathological prion protein is removed by using the filter according to the present invention at room temperature or under cooling for 72 hours, more preferably within 24 hours, particularly preferably within 12 hours, most preferably within 8 hours after collection of whole blood, which is then stored at room temperature or under refrigeration. In the case of removal of the pathological prion protein after storage with the purpose of obtaining a product of whole blood that does not contain pathological prion protein, pathological prion protein is removed by means of filter preferably 24 hours before use. In the present invention it is possible to prepare a product of whole blood that does not contain pathological prion protein and leukocytes.

(Product red blood cells that do not contain pathological prion protein)

To SOBR is authorized whole blood add a solution of preservative or anticoagulant such as CPD, CPDA-1, CP2D, ACD-A, ACD-B or heparin. Examples of the method of successive division into components include a method involving the removal of abnormal prion protein from whole blood, followed by centrifugation; and a method involving centrifugation of whole blood with subsequent removal of the pathological prion protein from red blood cells or erythrocytes and leucocytes film (indicated below as "BC").

When the centrifugation is carried out after removal of the pathological prion protein from a whole blood product red blood cells that do not contain pathological prion protein, is obtained by obtaining the product of whole blood that does not contain pathological prion protein similar to the product of whole blood that does not contain pathological prion protein, centrifugation product whole blood that does not contain pathological prion protein, and collecting the concentrated red blood cells in the lower level.

When whole blood is centrifuged to remove the pathological prion protein, you can apply two variants of centrifugation conditions: mild conditions centrifugation to separate the blood into red blood cells and platelet-rich plasma (PRP) and the hard conditions of centrifugation for the separation of blood into red blood cells, BC and platelet-poor plasma (PPP). PR is necessary, add a solution of a preservative, such as saline adenine, glucose and mannitol (SAGM), additive solution-1 (AS-1), additive solution 3 (AS-3), an additive solution-5 (AS-5), or mannitol-adenine-phosphate (MAP) to erythrocytes separated and selected from whole blood or erythrocytes containing BC, and product red blood cells can be filtered using the filter according to the present invention to obtain thereby the product of red blood cells that do not contain pathological prion protein.

Upon receipt of the product red blood cells that do not contain pathological prion protein, centrifugation is carried out preferably within 72 hours, preferably 48 hours, particularly preferably 24 hours, most preferably 12 hours after collection of whole blood at room temperature or under refrigeration. In case of pathological prion protein is removed before storage, the product is whole blood that does not contain pathological prion protein is prepared by removal of pathological prion protein by means of the filter preferably within 120 hours, more preferably 72 hours, particularly preferably 24 hours, most preferably 12 hours after blood sampling from a product of red blood cells stored at room temperature or under refrigeration. In case of pathological prion protein remove the Le storage, pathological prion protein is removed from the product red blood cells stored at room temperature or under cooling, preferably within 24 hours before use, thus obtaining a product of red blood cells that do not contain pathological prion protein.

(Getting the product platelets that do not contain pathological prion protein)

To the collected whole blood add the preservative solution of anticoagulant, such as CPD, CPDA-1, CP2D, ACD-A, ACD-B or heparin.

Examples of the method of successive division into components include: a method involving the removal of abnormal prion protein from whole blood, followed by centrifugation; and a method involving centrifugation of whole blood with subsequent removal of the pathological prion protein from PRP or platelet.

When the centrifugation is carried out after the removal of abnormal prion protein from a whole blood product platelets that do not contain pathological prion protein, is obtained by obtaining the product of whole blood that does not contain pathological prion protein, in the same way as when receiving product whole blood that does not contain pathological prion protein, centrifugation of whole blood that does not contain pathological prion protein, PRP collection in the upper layer, centrif the responses of the upper layer and collecting the concentrated platelets (PC) in the lower layer.

When whole blood is centrifuged to remove the pathological prion protein, you can apply two variants of centrifugation conditions: mild conditions centrifugation to separate the blood into red blood cells and PRP and hard conditions of centrifugation for the separation of blood into red blood cells, BC and PPP. In the case of soft conditions centrifugation product platelets that do not contain pathological prion protein, obtained by filtering PRP separated from whole blood using a filter for removal of pathological prion protein, repeated centrifugation of the filtrate and collect the PC in the lower level, or by centrifugation of the PRP for separating platelets from PPP and filtering PC in the lower level with the use of the filter for removal of pathological prion protein. In the case of the hard conditions of centrifugation product platelets that do not contain pathological prion protein prepared by adding, if necessary, the solution of the preservative or plasma to BC, separated from whole blood and collected in the amount of from one to several tens units, centrifugation of the solution, collecting the upper layer for receiving the concentrated platelets and filtration of platelets using a filter for removal of pathological prion protein.

Upon receipt of the product platelets, the e containing pathological prion protein, whole blood stored at room temperature, collected within 24 hours, more preferably 12 hours, particularly preferably 8 hours after collection of whole blood. In the case of removal of the pathological prion protein product of whole blood that does not contain pathological prion protein is prepared by removal of pathological prion protein by using the filter preferably within 120 hours, more preferably 78 hours, particularly preferably 24 hours, most preferably 12 hours after collection product platelets, which are then stored at room temperature. In case of pathological prion protein is removed after storage, the pathological prion protein can be removed by using the filter from the product platelets stored at room temperature or under cooling, for 24 hours before use, thus obtaining the product platelets that do not contain pathological prion protein.

(Obtaining plasma product that does not contain pathological prion protein)

To the collected whole blood add the preservative solution of anticoagulant, such as CPD, CPDA-1, CP2D, ACD-A, ACD-B or heparin.

Examples of the method of successive division into components include: a method involving the removal of abnormal prion protein from whole blood to follow them by centrifugation; and method including centrifugation of whole blood with subsequent removal of the pathological prion protein from PPP or PRP.

When the centrifugation is carried out after removal of the pathological prion protein from whole blood, plasma product that does not contain leukocytes, obtained by obtaining the product of whole blood that does not contain pathological prion protein, in the same way as when receiving product whole blood that does not contain pathological prion protein, centrifugation of whole blood that does not contain pathological prion protein and collection of plasma in the upper layer.

When whole blood is centrifuged to remove the pathological prion protein, you can apply two variants of centrifugation conditions: mild conditions centrifugation to separate the blood into red blood cells and PRP and hard conditions of centrifugation for the separation of blood into red blood cells, BC and PPP. In the case of soft conditions centrifugation the plasma product that does not contain pathological prion protein, obtained by filtering PRP using a filter for removal of pathological prion protein, after filtering PRP, repeated centrifugation of the filtrate and collect the PPP in the supernatant, or by separation of PRP on PPP and platelets by centrifugation and filtration PPP using the filter at the Alenia pathological prion protein. In the case of the hard conditions of centrifugation the plasma product that does not contain pathological prion protein is prepared by PPP filtering using a filter for removal of pathological prion protein.

Upon receipt of the plasma product that does not contain pathological prion protein, whole blood stored at room temperature or cooled, centrifuged, preferably within 72 hours, preferably 48 hours, particularly preferably 24 hours, most preferably 12 hours after collection of whole blood. Alternatively, a plasma product that does not contain pathological prion protein is prepared by removal of pathological prion protein by using the filter preferably within 120 hours, more preferably 72 hours, particularly preferably 24 hours, most preferably 12 hours after collection of the blood plasma product, which is then stored at room temperature or under refrigeration. In case of pathological prion protein is removed after storage, the pathological prion protein can be removed by using the filter from the product of the plasma stored at room temperature or under cooling, preferably within 24 hours before use, thus generating a plasma product that does not contain pathological prion b is the left main coronary artery.

The term "filter for removal of leucocytes in the present invention means a filter capable of removing leukocytes by filtration of the blood product at the level of 99% or more, preferably of 99.9% or more, more preferably of 99.99% or more. When the filter efficiency is expressed as the efficiency of removal of leukocytes, the filter shows the value calculated by the following formula (1). Thus, the filter has a removal efficiency of cells equal to 2 or more, preferably 3 or more, more preferably 4 or more.

The removal efficiency of leukocytes = -Log10{[concentration of cells (cells per µl) in the blood after filtration]/ [concentration of cells (cells per µl) of blood to filter]} (1).

The sterilization method of the filter according to the present invention includes: gas sterilization with ethylene oxide; sterilization by irradiation, such as sterilization, γ-irradiation or electron beam; and sterilization by autoclaving. Preferred sterilization by irradiation or autoclaving. To achieve high efficiency of removal of leukocytes from whole blood filter is preferably subjected to as sterilization by irradiation and sterilization by autoclaving. Between sterilization by irradiation and sterilization by autoclaving is not priority one is to more preferably, to sterilization by autoclaving was carried out prior to sterilization by irradiation. Use media that is coated with polymer, subjected to sterilization by autoclaving may reduce the number of pathological prion protein, adsorbiruyuschee on the carrier coated with the polymer, the product of whole blood. Perhaps this is due to the competitive adsorption of proteins in the product whole blood, because the sterilization by autoclaving does not cause any changes in the chemical properties of the polymer on the surface of the carrier. On the other hand, in the case of sterilization by irradiation with high removal efficiency of prion can be achieved even in the product whole blood. The reason for this is unclear, but it can be assumed that the sterilization filter irradiation causes changes in the chemical properties of the polymer on the surface of the carrier, which leads to the preferred charge and balance the composition of the three elements of polymer adsorption pathological prion protein. However, it is noteworthy that the authors of the present invention found that a high removal efficiency of pathological prion protein from whole blood can be achieved even after sterilization by autoclaving, provided that it is performed after sterilization by irradiation. It is expected that if sterilization by autoclaving spend the SJ after sterilization by irradiation, saved specificity of adsorption of pathological prion protein, which allows for high efficiency removal of pathological prion protein regardless of sterilization by autoclaving. In the case of using an embedded system, if the filter is pre-sterilized by irradiation, high efficiency removal of pathological prion protein can be achieved even after sterilization by autoclaving.

[Example]

The present invention is described in more detail in the examples, which are not intentionally limit the present invention.

The numerical values used in the examples and comparative examples were obtained in the following ways.

(The specific surface area of the filter material)

The term "specific surface area (m2/g)" in the present invention means the surface area per unit mass of non-woven material, a specific method of gas adsorption (method BET) using Accusorb 2100 (production Shimadzu Corp) or its equivalent. Specific surface area is determined as follows: a tube for samples fill 0.50 to 0.55 g of the carrier; from the tube to remove the air under reduced pressure of 1·10-4mm RT. senior (at room temperature) for 20 hours in the device Accusorb; adsorb krypton gas, which is known PLO the location area of adsorption as gas-adsorbate to the surface of the nonwoven material at a temperature equivalent to the temperature of liquid nitrogen; calculate the total surface area in the non-woven material based on the adsorbed amount; and divide the total surface area to the weight of the nonwoven material.

(Measurement of average fiber diameter)

Did electron microscopic pictures of five random points on a non-woven material. Each photo has put a transparent sheet with a printed grid. Measured the diameter of the thread at the point of intersection with the grid (n=100) and determined the diameter by converting the measured diameter, using as scale polystyrene latex with a known diameter.

(The amount of polymer per unit area of the total surface area of the material)

The term "total surface area (m2material" in the present invention means the value obtained by multiplying the mass of material (g) specific surface area (m2/g) of material. The amount of polymer (mg/m2) per unit area (m2) the total surface area of the material according to the present invention was determined by NMR analysis of the solution of a certain area (weight) of material dissolved in a deuterated solvent common to the carrier and covering agent. For example, the prescribed amount of material, including polyester non-woven material, the open polymer, containing methyl methacrylate, dimethylaminoethylmethacrylate and 2-hydroxynitriles, was dissolved in deuterated 1,1,1,3,3,3-hexafluoro-2-propanol. Determined the ratio of the signals is clearly related to the non-woven material (i.e. the protons on the benzene ring), and signals clearly related to the covering material (i.e. protons on the methyl group adjacent to the methyl methacrylate). It was further determined the amount of polymer per unit weight of the nonwoven material of relationship intensity and the copolymerization composition of coating material defined separately. The amount of polymer per unit weight of the nonwoven material can be converted into an amount of polymer on the total surface area of the material, using the specific surface area of the material that fills the filter.

(Analysis of blood filtering: Examples 1-13 and Comparative Examples 1-9)

To evaluate blood as blood products used products of red blood cells, each product was obtained in the following way: immediately after blood collection to 100 ml of blood was added 14 ml of CPD solution that serves as an anticoagulant; mixed mixture; keep the mixture in the refrigerator for 72 hours; the mixture was centrifuged; to the separated red blood cells containing BC, was added SAGM; and leave the mixture for 1 hour (below referred to as "unfiltered blood").

The filter used is La assessment, was obtained by filling a container of polyester non-woven material (average fiber diameter of 1.2 μm, the weight of the substrate per unit area of 40 g/m2the specific surface area of 1.47 m2/g)coated with the polymer obtained in each of examples and comparative examples (without coating in Comparative Example 6). Column (effective filtration area 1.3 cm2) fills eight covered by non-woven materials, and to the input of the column was connected to a syringe filled with blood before filtration through a vinyl chloride hose (inner diameter 3 mm, outer diameter 4.2 mm). Then the blood ran through the column with a flow rate 0,175 ml/min using a syringe pump in the refrigerator and collected in the amount of 3 ml (indicated below as "filtered blood"). Covered with non-woven materials (without coating in Comparative Example 6) were subjected to sterilization, γ-irradiation at 25 kGy in Examples 1-5 and 11 and Comparative Examples 1-9; sterilization electron beam at 30 kGy in Examples 8 and 12; sterilize by autoclaving at 115°C for 59 minutes in Examples 6, 7, 10 and 13; sterilization electron beam at 30 kGy and then sterilized by autoclaving at 115°C for 59 minutes in Example 9; and not subjected to sterilization in Comparative example 10.

[The removal efficiency of leukocytes]

The removal efficiency of lenoci the s was calculated by the above formula (1), based on the concentrations of leukocytes before and after filtering. The concentration of cells was measured by the method of flow cytometry (unit FASCcalibur, production Becton Dickinson) in 100 μl of each blood sample using the Leucocount kit, which includes beads (manufactured by Nippon Becton Dickinson Company, Ltd.).

[The pressure in the processing of blood]

The pressure in the processing blood was measured at the end of filtration using a pressure gauge connected with the hose on the inlet side of the column.

[The level of hemolysis in the blood after filtration]

The level of hemolysis in the blood after filtration was calculated based on the level of free hemoglobin in plasma in relation to the total level of hemoglobin in the product red blood cells after 42 days after filtering.

A more accurate method of calculating the level of hemolysis is as follows:

(1) the Content of total hemoglobin (g/DL) and hematocrit (%) in product red blood cells after filtration was determined using an automatic counter blood cells.

(2) Collected a sample of the product red blood cells after filtering in the amount of 2 ml and was centrifuged at 1750g for 10 minutes

(3) Measured values of the absorption of the supernatant by scanning a wavelength between 630 nm and 500 nm using a spectrophotometer.

(4) the Concentration of free hemoglobin in plasma was measured according the manual, described in Clin. Biochem. 8, 96-102 (1975).

(5) the Level of hemolysis was calculated according to the following formula (2).

The level of hemolysis (%)=(100-level hematocrit (%))·(concentration of free hemoglobin (g/DL))/concentration of total hemoglobin (g/DL) (2).

(Evaluation of the effectiveness of the removal of the pathological prion protein: Examples 1-13 and Comparative examples 1, 3, 5-8, and 10)

[Preparation of filter]

Polyester non-woven material P (the average fiber diameter of 12 μm, the weight of the substrate per unit area of 30 g/m2the specific surface area of 0.24 m2/g), polyester nonwoven fabric (average fiber diameter of 2.5 μm, the weight of the substrate per unit area of 60 g/m2the specific surface area of 0.8 m2/g), polyester non-woven material B (average fiber diameter of 1.8 μm, the weight of the substrate per unit area of 60 g/m2the specific surface area of 1.1 m2/g), and polyester non-woven material C (average fiber diameter of 1.2 μm, the weight of the substrate per unit area of 40 g/m2the specific surface area of 1.47 m2/g)coated with the polymer produced in each of examples and comparative examples (without coating in comparative example 6), was used as the filter material. Filter materials P, A, B and C were formed in the order of P-A-B-C with the input side, and then from the output side was formed B' (PI is trouse material, identical In), A' (filter material that is identical to A), and P' (filter material which is identical to P), thus receiving filter having a symmetrical structure P-A-B-C-B'-A'-P'. The obtained filter material is made between sheets of flexible vinyl chloride resin with ports serving for entry and exit of blood, peripheral border between the filtering material and a flexible sheet glued and joined by means of high frequency welding machines, thus providing the filter with an effective filtration area 56 cm2.

The resulting filters were subjected to sterilization, γ-irradiation at 25 kGy in Examples 1-5 and 11 and Comparative Examples 1-9; sterilization electron beam at 30 kGy in Examples 8 and 12; sterilize by autoclaving at 115°C for 59 minutes in Examples 6, 7, 10 and 13; sterilization electron beam at 30 kGy and then sterilized by autoclaving at 115°C for 59 minutes in Example 9; and not subjected to sterilization in Comparative example 10.

[The hamster brain infected with scrapie]

Pathological prion protein Sc237 isolated from passage hamster 263K for the development of scrape, instilled hamster, and after 65-70 days the hamster brain was isolated and used as hamster brain infected with scrapie.

[Homogenate]

The brains of hamsters infected scrape, Sonicare is whether the sucrose solution with a concentration of 320 mm to obtain a homogenate with the mass-volume concentration (g/100 ml solution), equal to 10% (hereinafter indicated as wt./vol.%). The homogenate was centrifuged at 4°C and 80g within 1 minute and was used as a homogenate to add (hereinafter abbreviated to "homogenate").

[Microsomal fraction]

The homogenate was centrifuged at 5000g for 20 minutes, the supernatant further centrifuged at 100000g for 1 hour. The obtained precipitation resuspendable in solution (150 mm NaCl, 20 Tris-HCl, pH 7,4), the suspension was centrifuged at 100000g for 1 hour. The obtained precipitation resuspendable in PBS pH 7,4 (saline phosphate buffer) to obtain the microsomal fraction to add (below abbreviated to "microsomal fraction").

[Method 1 for determining the concentration of protease To: Examples 1, 2, 4 and 13]

(Sample not treated with protease K)

50 μl of Homogenate and 50 μl of the product of erythrocytes containing the homogenate was mixed separately, each with 450 μl of the sample buffer, the mixture was incubated at 100°C±5°C for 5-10 minutes or at 70-80°C for 10-15 minutes.

(Sample treated with protease K)

50 μl of Homogenate and 50 μl of the product of erythrocytes containing the homogenate was mixed separately with solutions containing protease in different concentrations and 0.6-1% sarkosyl, mixtures were incubated at 37°C±4°C for 1 hour ± 5 minutes. To the mixtures were added to 300 μl of the sample buffer and the mixture is incubated separately at 100°C±5°C for 5-10 minutes or at 70-80°C for 10-15 minutes.

Samples not treated with protease K and treated with protease K was analyzed using Western blot analysis to determine the minimum concentrations of the protease To to proteins capable of nonspecific react with the antibody, and normal prion protein was fully degradibility protease K and not detected, while the pathological prion protein detected.

[Method 2 determine the concentration of the protease To Examples 3 and 6 and Comparative examples 5, 6 and 10]

(Sample not treated with protease K)

5 ml of blood containing the microsomal fraction, and 5 ml of blood containing normal prion protein, centrifuged separately at 4000g for 20 minutes. After that, 3 ml of supernatant was centrifuged at 100000g and 4°C±2°C for 1 hour. The obtained precipitation resuspendable in 100 ál of sample buffer and incubated suspension at 100°C±5°C for 5-10 minutes.

(Sample treated with protease K)

5 ml of blood containing the microsomal fraction, and 5 ml of blood containing normal prion, centrifuged separately at 4000g for 20 minutes. Solutions with different concentrations of protease K was mixed with 3 ml of supernatant, and the mixture was centrifuged at 100000g and 4°C±2°C for 1 hour. The obtained precipitation resuspendable in 100 μl of buffer DL the samples, and the suspension incubated at 100°C±5°C for 5-10 minutes.

Samples not treated with protease K, and the samples treated with protease K was analyzed using Western blot analysis to determine the minimum concentrations of the protease To to proteins capable of nonspecific react with the antibody, and normal prion protein was fully degradibility protease K and not detected, while the pathological prion protein detected.

[Method 3 determination of the concentration of the protease To: Example 5 and Comparative Examples 1, 3, 7, and 8]

(Sample not treated with protease K)

50 μl of Plasma, containing the microsomal fraction, and 50 μl of plasma separately mixed with 50 μl of the sample buffer and the mixture incubated at 100°C±5°C for 5-10 minutes.

(Sample treated with protease K)

Solutions with different concentrations of protease K was mixed with 3 ml of plasma containing the homogenate obtained from hamster brain infected with scrapie, and 3 ml of plasma containing the microsomal fraction obtained from hamster brain infected with scrapie, and the mixture was centrifuged at 20000g and 4°C±2°C for 1 hour. After centrifugation to the obtained precipitation was added to 100 ál of sample buffer and kept at 100°C±5°C for 5-10 minutes to complete the reaction.

Samples not treated with protease K,and samples treated with protease K was analyzed using Western blot analysis to determine the minimum concentrations of the protease To to proteins capable of nonspecific react with the antibody, and normal prion fully degradibility protease K and not detected, while the pathological prion protein detected.

[Method 4 to determine the concentration of protease To: Examples 7-9]

Product whole blood after removal of cells was centrifuged at 4000g for 30 minutes, and to 11.7 ml of the obtained supernatant was added 1.3 ml of microsomal fractions (indicated below as "unfiltered blood with additives"), while to 12 ml of the supernatant did not add any substances (below labeled "unfiltered blood without additives").

(Sample not treated with protease K)

3 ml of Unfiltered blood with additives and 3 ml of unfiltered blood without additives was centrifuged separately at 100000g and 4°C±2°C for 1 hour. The obtained precipitation resuspendable in 100 µl of sample buffer, and the suspension was incubated at 100°C±5°C for 5-10 minutes.

(Sample treated with protease K)

1-2% (wt./about) Sarkosyl and solutions protease To different concentrations was mixed with 3 ml of unfiltered blood with additives or 3 ml of unfiltered blood without additives, and the mixture was centrifuged at 100000g n and 4°C±2°C for 1 hour. The obtained precipitation resuspendable in 100 µl of sample buffer, and the suspension was incubated separately at 100°C±5°C for 5-10 minutes.

Samples not treated with protease K, and the samples treated with protease K was analyzed using Western blot analysis to determine the minimum concentrations of the protease To to proteins capable of nonspecific react with the antibody, and normal prion protein is not detected due to treatment with protease K, while the pathological prion protein detected.

[Method 5 for determining the concentration of protease To: Examples 10-12]

Product red blood cells after removal of cells was centrifuged at 4000g for 30 minutes, and to 11.7 ml of the obtained supernatant was added 1.3 ml of microsomal fractions (indicated below as "unfiltered blood with additives"), while to 12 ml of the supernatant did not add any substances (below labeled "unfiltered blood without additives").

(Sample not treated with protease K)

3 ml of Unfiltered blood with additives and 3 ml of unfiltered blood without additives was centrifuged separately at 100000g and 4°C±2°C for 1 hour. The obtained precipitation resuspendable in 100 µl of sample buffer, and the suspension was incubated at 100°C±5°C for 5-10 minutes.

(Sample treated with protease K)

1-2% (wt./about) With rasila and solutions protease To different concentrations was mixed with 3 ml of unfiltered blood with additives or 3 ml of unfiltered blood without additives, and the mixture was centrifuged at 100000g at 4°C±2°C for 1 hour. The obtained precipitation resuspendable in 100 µl of sample buffer, and the suspension was incubated separately at 100°C±5°C for 5-10 minutes.

Samples not treated with protease K, and the samples treated with protease K was analyzed using Western blot analysis to determine the minimum concentrations of the protease To to proteins capable of nonspecific react with the antibody, and normal prion protein is not detected due to treatment with protease K, while the pathological prion protein detected.

[Test 1 the efficiency of removal of pathological prion protein: Examples 1, 2, 4 and 13]

Bought and used the product red blood cells, obtained according to the European standard. The homogenate was added to the product red blood cells at a rate of 13 ml relative to 1 unit of product red blood cells at room temperature, thus obtaining unfiltered blood with additives. The resulting solution was filtered using a filter that includes a polyester nonwoven material coated with the polymers obtained in Examples 1, 2, 4 and 13 by means of gravity from a height of 100 cm, and the blood was collected, thus obtaining filtered blood with additives. First unfiltered blood with additives and filtered blood with additives races is varali in cell lysate (obtained by dissolution of 11.45 g of ammonium oxalate, 433 mg of potassium dihydrophosphate and 567 mg monohydrogenphosphate sodium in 1 liter of distilled water for injection). As unfiltered blood with additives, and the filtered blood with additives were mixed with the cell lysate in a volume ratio of 1:3 and the mixture was dissolved at room temperature for 30 minutes. After dissolution was performed three times sonicate each time for 15±5 seconds when the power level of 50±10%) intermittently for 30±10 sec. To unfiltered blood with additives and filtered blood with additives were added to the solution of the protease To the concentration determined by the method 1 for determining the concentration of protease K, and 0.6-1% sarkosyl, cleavage by protease K was carried out at 37°C±3°C for 1 hour ±5 minutes, and then stopped by adding 10 mm Pefabloc. Next, the resulting mixture was centrifuged at 100000g and 4°C±2°C for 1 hour, was added to 500 ál of sample buffer to the obtained precipitation, and then incubated at 100°C±5°C for 5-10 minutes or at 70-80°C for 10-15 minutes.

[Test 2 the efficiency of removal of pathological prion protein: example 3 and comparative examples 5 and 6]

Bought and used the product red blood cells that do not contain leukocytes, manufactured according to European standard. The product was kept for 1-2 days in the refrigerator at 4°C, was added 12 ml of microsomal fraction to 1 unit of product eritria is s, does not contain leukocytes, at room temperature, thus obtaining unfiltered blood with additives. The resulting solution was filtered using a filter that includes a polyester nonwoven material coated with the polymer obtained in Example 3 and Comparative Examples 5 and 6 (not covered by the polymer in Comparative Example 6) using the force of gravity from a height of 100 cm, and the blood was collected, thus obtaining filtered blood with additives. Unfiltered blood with additives and filtered blood with additives was centrifuged at 4000g for 30 minutes, supernatant was divided into aliquots. To supernatants added protease in a concentration of 1000 μg/ml, and the mixture was centrifuged at 100000g and 4°C±2°C for 1 hour. After centrifugation, precipitation resuspendable in 100 µl of sample buffer, and the suspension was incubated at 100°C±5°C for 5-10 minutes.

[Test 3 the efficiency of removal of pathological prion protein: Example 5 and Comparative Examples 1, 3, 7, and 8]

Acquired fresh frozen plasma, manufactured according to European standard. On the day of the filtered plasma was thawed at 37°C and was added to the microsomal fraction in the volume of 8.6% compared to the mass of the plasma, thus receiving unfiltered blood with additives. Unfiltered blood with additives (150±2 g) was filtered using a filter that includes polyester non-woven material, covered by the polymer obtained in Example 5 and Comparative Examples 1, 3, 7 and 8 using the force of gravity from a height of 100 cm, and the blood was collected, thus obtaining filtered blood with additives. To 100 μl of unfiltered blood with additives and filtered blood with additives added protease K at a concentration of 600 μg/ml, the mixture was centrifuged at 20000g and 4°C±2°C for 1 hour. Precipitation resuspendable in 100 µl of sample buffer, and the suspension was incubated at 100°C±5°C for 5-10 minutes.

[Test 4 the efficiency of removal of pathological prion protein: Example 6 and Comparative Example 10]

Bought and used the product whole blood containing leukocytes, manufactured according to European standard. Thus obtained product whole blood was stored for one day in a refrigerator at 4°C, and 1 unit of product whole blood containing leukocytes was added 28 ml of microsomal fractions, thus receiving unfiltered blood with additives. The resulting solution was filtered using a filter that includes polyester non-woven material coated with the polymer obtained in Example 6, and the filter used in Comparative Example 10, using the force of gravity from a height of 100 cm, and the blood was collected, thus obtaining filtered blood with additives. Unfiltered blood to add the AMI and filtered blood with additives was centrifuged at 4000g for 30 minutes, supernatant was divided into aliquots. To supernatants added protease in a concentration of 1000 μg/ml, and the mixture was centrifuged at 100000g and 4°C±2°C for 1 hour. After centrifugation, precipitation resuspendable in 100 µl of sample buffer, and the suspension was incubated at 100°C±5°C for 5-10 minutes.

[Test 5 the efficiency of removal of pathological prion protein: Examples 7, 8, and 9]

Bought and used the product whole blood containing leukocytes, manufactured according to European standard. The product was stored for 1-3 days in the refrigerator at 4°C, and three units of product red blood cells were placed in a single package for blood. Next was adding 84 ml microsomal fractions at room temperature, thus obtaining unfiltered blood with additives. Unfiltered blood with additives was divided into three parts, and the resulting unfiltered blood with additives was filtered using a filter that includes a polyester nonwoven material coated with the polymer obtained in each of Examples 7, 8 and 9, by force of gravity from a height of 100 cm, and the blood was collected, thus obtaining filtered blood with additives. Unfiltered blood with additives and filtered blood with additives was centrifuged at 4000g for 30 minutes, supernatant was divided into aliquots. To supernatants added protease K To the end of the ation 25 u/ml, and the mixture was centrifuged at 100000g and 4°C±2°C for 1 hour. After centrifugation, precipitation resuspendable in 100 µl of sample buffer, and the suspension was incubated at 100°C±5°C for 5-10 minutes.

[Test 6 the efficiency of removal of pathological prion protein: Examples 10, 11 and 12]

Bought and used the product red blood cells that do not contain leukocytes, manufactured according to European standards. The product was stored for 2-4 days in the refrigerator at 4°C, and 1 unit of product red blood cells that do not contain leukocytes, was added to 9.5 ml of microsomal fractions at room temperature, thus obtaining unfiltered blood with additives. The resulting solution was filtered using a filter that includes a polyester nonwoven material coated with the polymer obtained in each of Examples 10, 11 and 12 and by force of gravity from a height of 100 cm, and the blood was collected, thus obtaining filtered blood with additives. Unfiltered blood with additives and filtered blood with additives was centrifuged at 4000g for 30 minutes, supernatant was divided into aliquots. To supernatants added protease K To a concentration of 25 u/ml, and the mixture was centrifuged at 100000g and 4°C±2°C for 1 hour. After centrifugation, precipitation resuspendable in 100 µl of sample buffer, and the suspension was incubated at 100°C±5°is within 5-10 minutes.

[Analysis of the title of the pathological prion protein]

Blood products unfiltered blood with additives and filtered blood with additives were added to the sample buffer, the mixture was heated and it was further revealed pathological prion protein using a standard method, Western blotting, using as the first antibody antibody 3F4, specifically bind to the prion protein. Analysis of all products unfiltered blood with additives and filtered blood with additives was repeated several times in a series of continuous three-fold dilutions, ED50was determined by Spearman (Brit. J. Of Psychology 1908; 2: 227 ff.) and Kaerber (Naunyn Schmiedeberg''s Arch. Exp. Path. Pharmak. 1931; 152: 380 ff.). Next, all values led to the ED50per unit volume (1 ml), the logarithm of this value was taken as the titer. The method of calculation of the ED50below.

[Formula 1]

Y0positive exponent of the highest dilution of the sample showing positive test results in all parallel dilutions;

d is the logarithm of the step cultivation;

ΣYi- the sum of the percentage of samples where the pathological prion protein was detected at a dilution of sample in Y0times or more.

[The removal efficiency of pathological prion protein]

The removal efficiency of pathological prion protein was calculated from etousa the formula (4).

The removal efficiency of pathological prion protein = titer of pathological prion protein in unfiltered blood with additives titer of pathological prion protein in the filtered blood with additives (4)

[Example 1]

The polymerization was performed by adding dropwise of a solution of ethanol obtained by dissolving the polymerized monomers and the initiator diazo-compounds in ethanol used as a solvent for polymerization with stirring at 78°C under nitrogen atmosphere.

Charged curable monomers include 20 molar percent of methyl methacrylate (below referred to as "MMA"), used as hydrophobic polymerized monomer, 5 molar percent of dimethylaminoethylmethacrylate (indicated below as "DM"), used as a polymerized monomer containing a basic nitrogen-containing part, 75 molar percent of 2-hydroxyethylmethacrylate (indicated below as "HEMA"), is used as the monomer containing a protonic neutral hydrophilic part. The polymer solution was purified by using an excess of water and dried under reduced pressure. The copolymerization composition of the polymer was analyzed using 1H-NMR. The results practically coincide with the composition of the charged polymerized monomer, copolymerization compositions MA, DM and HEMA in the polymer equal to 20 molar percent, 5 molar percent and 75 molar percent, respectively. Average molecular weight (Mw) was 210000. As the coating solvent used ethanol was used, the polymer concentration of 0.8 wt./vol.%. The polymer solution in the above concentrations were covered with polyester non-woven material used as the material. Covered the number was 21 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by method 1 was tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein was 4,12, of 9.8 kPa, 0.2% and 2.0 or more, respectively.

[Example 2]

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 20 molar percent MMA, 13 molar percent DM and 67 mole% HEMA. The copolymerization composition of the polymer was analyzed using 1H-NMR. Composition copolymerization of MMA, DM and HEMA in the polymer was 20 molar percent, 13 molar percent and 67 molar numbers of the respectively. Average molecular weight (Mw) was $ 250000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material Covered With. the number was 22 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by method 1 was tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein was 5,68, 8,1 kPa, 0.5% and 2.0 or more, respectively.

[Example 3]

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 30 molar percent MMA, 10 molar percent DM and 60 molar percent HEMA. The copolymerization composition of the polymer was analyzed using 1H-NMR. Composition copolymerization of MMA, DM and HEMA in the polymer was 30 molar percent, 10 molar percent and 60 molar percent, respectively. Average molecular weight (Mw) was 210000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven mother of the L. S. Covered the number was 20 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by method 2 were tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein was 5,68, 11,4 kPa, 0.4 percent and 3.8, respectively.

[Example 4]

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 40 molar percent MMA, 5 molar percent DM and 55 molar percent HEMA. The copolymerization composition of the polymer was analyzed using 1H-NMR. Composition copolymerization of MMA, DM and HEMA in the polymer was 40 molar percent, 5 molar percent and 55 molar%, respectively. Average molecular weight (Mw) was 170000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material Covered With. the number was 22 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filter is AI and the removal efficiency of pathological prion protein by method 1 was tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein amounted to 5.17, 12,5 kPa, 0.3% and 2.0 or more, respectively.

[Example 5]

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 40 molar percent MMA, 13 molar percent DM and 47 mole% HEMA. The copolymerization composition of the polymer was analyzed using 1H-NMR. Composition copolymerization of MMA, DM and HEMA in the polymer was 40 molar percent, 13 molar percent and 47 mole percent, respectively. Average molecular weight (Mw) was 180000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material Covered With. the number was 22 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by method 3 was tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein was 5,68, 10,9 kPa, 0.5% and 2.4 matched with the public.

[Example 6]

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 30 molar percent MMA, 10 molar percent DM and 60 molar percent HEMA. The copolymerization composition of the polymer was analyzed using 1H-NMR. Composition copolymerization of MMA, DM and HEMA in the polymer was 30 molar percent, 10 molar percent and 60 molar percent, respectively. Average molecular weight (Mw) was 210000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material Covered With. the number was 20 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by the method 4 was tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein was 5,86, to 13.2 kPa, 0.4 percent and 1.5, respectively.

[Example 7]

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 30 molar percent MMA, 10 molar percent DM and 60 molar the percent HEMA. The copolymerization composition of the polymer was analyzed using 1H-NMR. Composition copolymerization of MMA, DM and HEMA in the polymer was 30 molar percent, 10 molar percent and 60 molar percent, respectively. Average molecular weight (Mw) was 210000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material Covered With. the number was 20 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by the method 5 was tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein was 5,12, of 11.5 kPa, 0.3 percent and 1.2, respectively.

[Example 8]

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 30 molar percent MMA, 10 molar percent DM and 60 molar percent HEMA. The copolymerization composition of the polymer was analyzed using 1H-NMR. Composition copolymerization of MMA, DM and HEMA in the polymer was 30 molar percent, 10 molar percent and 60 molar percent of the respectively. Average molecular weight (Mw) was 210000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material Covered With. the number was 20 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by the method 5 was tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein was 4,68, 12.0 kPa, 0.3 per cent and 1.6, respectively.

[Example 9]

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 30 molar percent MMA, 10 molar percent DM and 60 molar percent HEMA. The copolymerization composition of the polymer was analyzed using 1H-NMR. Composition copolymerization of MMA, DM and HEMA in the polymer was 30 molar percent, 10 molar percent and 60 molar percent, respectively. Average molecular weight (Mw) was 210000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material C. Pokr is the same number was 20 mg/m 2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by the method 5 was tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein was 5,43, 11,8 kPa, 0.2 per cent and 1.7, respectively.

[Example 10]

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 30 molar percent ethyl methacrylate (indicated below as "EMA"), 10 molar percent DM and 60 molar percent HEMA. The copolymerization composition of the polymer was analyzed using 1H-NMR. The copolymerization composition EMA, DM and HEMA in the polymer was 30 molar percent, 10 molar percent and 60 molar percent, respectively. Average molecular weight (Mw) was 220000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material Covered With. the number was 19 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after Phi is Tracie and the removal efficiency of pathological prion protein by method 6 was tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein was 5,22, to 10.5 kPa, 0.3% and 4,1 or more, respectively.

[Example 11]

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 30 molar percent MMA, 10 molar percent of diethylaminoethylmethacrylate (hereinafter referred to as "DE") and 60 molar percent HEMA. The copolymerization composition of the polymer was analyzed using 1H-NMR. The copolymerization composition EMA, DM and HEMA in the polymer was 30 molar percent, 10 molar percent and 60 molar percent, respectively. Average molecular weight (Mw) was 200000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material Covered With. the number was 20 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by method 6 was tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathologizes the th prion protein was 4,58, to 12.3 kPa, 0.4% and 4,1 or more, respectively.

[Example 12]

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 30 molar percent MMA, 10 molar percent DM and 60 molar percent of hydroxypropylmethacrylate (hereinafter referred to as "HPMA"). The copolymerization composition of the polymer was analyzed using 1H-NMR. Composition copolymerization of MMA, DM and HPMA in the polymer was 30 molar percent, 10 molar percent and 60 molar percent, respectively. Average molecular weight (Mw) was 280000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material Covered With. the number was 22 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by method 6 was tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein was 4.83, 15,0 kPa, 0.2% and 4,1 or more, respectively.

[Example 13]

Polymerization, purification, and drying was carried out similarly to Example 1, excluding the rising use of relations charge of monomers 40 molar percent of butyl acrylate (hereinafter referred to as "BA"), used as hydrophobic polymerized monomer, 10 molar percent of diethylaminoethylmethacrylate (hereinafter referred to as "DEA"), used as a polymerized monomer containing a basic nitrogen-containing part, and 50 molar percent of hydroxyethylacrylate (hereinafter referred to as "HBA")used as a monomer containing a protonic neutral hydrophilic part. The copolymerization composition of the polymer was analyzed using 1H-NMR. The copolymerization composition BA, DEA and HPA in the polymer was 40 molar percent, 10 molar percent and 50 molar percent, respectively. Average molecular weight (Mw) was 150000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material Covered With. the number was 21 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by method 1 was tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein comprised of 4.75, and 11.5 kPa, 0.3% and 2.0 or more according to the respectively.

[Comparative Example 1]

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 30 molar percent MMA, 3 molar percent DM and 67 mole% HEMA. The copolymerization composition of the polymer was analyzed using 1H-NMR. Composition copolymerization of MMA, DM and HEMA in the polymer was 30 molar percent, 3 molar percent and 67 mole percent, respectively. Average molecular weight (Mw) was 200000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material Covered With. the number was 19 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by method 3 was tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein was 4,88, 10,2 kPa, 0.2 per cent and 0.5, respectively. When the composition of the polymerization, the polymerized monomer containing a basic nitrogen-containing part, was less than 5 molar percent, decreased the adsorption pathologic the ski prion protein, that led to a decrease in the efficiency of removal of pathological prion protein.

[Comparative example 2]

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 30 molar percent MMA, 16 molar percent DM and 54 mole% HEMA. The copolymerization composition of the polymer was analyzed using 1H-NMR. Composition copolymerization of MMA, DM and HEMA in the polymer was 30 molar percent, 16 molar percent and 54 mole percent, respectively. Average molecular weight (Mw) was $ 190000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material Covered With. the number was 21 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection and the level of hemolysis after filtration was tested in the ways described. The pressure during the processing of the blood and the level of hemolysis was 11.8 kPa, 2.3 per cent respectively. In the case of filtering by cooling with the increase in the copolymerization composition of the polymerized monomer containing a basic nitrogen-containing part, hemolysis was observed, so the level of hemolysis did not meet the standard (0,8%).

[Comparative Example 3]

Poly is anizatio, cleaning and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 15 molar percent MMA, 10 molar percent DM and 75 molar percent HEMA. The copolymerization composition of the polymer was analyzed using 1H-NMR. Composition copolymerization of MMA, DM and HEMA in the polymer was 15 molar percent, 10 molar percent and 75 molar percent, respectively. Average molecular weight (Mw) was 220000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material Covered With. the number was 20 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by method 3 was tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein was 5,11, 7,8 kPa, 0.3 per cent and 0.5, respectively.

[Comparative Example 4]

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 45 molar percent MMA, 10 molar percent DM and 45 molar percent HEMA. To the position copolymerization polymer was analyzed using 1H-NMR. Composition copolymerization of MMA, DM and HEMA in the polymer was 45 molar percent, 10 molar percent and 45 molar percent, respectively. Average molecular weight (Mw) was 180000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material Covered With. the number was 22 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection and the level of hemolysis after filtration was tested in the ways described. However, the test was discontinued because of the low fluidity of the blood product, a high pressure during treatment (more than 60 kPa) and fear of breakage of the hose and syringe, so that the measurement of the efficiency of removal of leukocytes and level of hemolysis was not possible.

[Comparative Example 5]

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 3 molar percent DM and 97 mole% HEMA. The copolymerization composition of the polymer was analyzed using 1H-NMR. The copolymerization composition DM and HEMA in the polymer was 3 molar percent and 97 molar%, respectively. Average molecular weight (Mw) was of 550,000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, polzuyuschimsya as covering the solvent, covered polyester non-woven material With a specific surface area of 1.47 m2/g, average diameter of the fibers of 1.2 μm and a mass per unit area of 40 g/m2. Covered the number was 7 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by method 2 were tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein amounted to 4.92, to 6.8 kPa, 0.2 per cent and 0.2, respectively.

[Comparative Example 6]

Used polyester non-woven material With a specific surface area of 1.47 m2/g, an average fiber diameter of 1.2 μm and a mass per unit area of 40 g/m2without the polymer coating. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by method 2 were tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein was 4,55, 20,3 kPa, 0.2% and 0,0 respectively.

[Comparative is an Example of 7]

As the acidic monomer used methacrylic acid containing a carboxyl group (hereinafter denoted by "MAA").

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 30 molar percent MMA, 10 molar percent of the MAA and 60 molar percent HEMA. The copolymerization composition of the polymer was analyzed using 1H-NMR. Composition copolymerization of MMA, MAA and HEMA in the polymer was 30 molar percent, 10 molar percent and 60 molar percent, respectively. Average molecular weight (Mw) was $ 230000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material Covered With. the number was 22 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by method 3 was tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein was are 5.36, of 10.0 kPa, 0.4% and 0,0 respectively. The polymer containing acidic monomer, proved ineffective against removal the pathological prion protein.

[Comparative example 8]

Polymerization, purification, and drying was carried out similarly to Example 1, except for using the relationship of the charge of monomers 10 molar percent DM and 90 molar percent HEMA. The copolymerization composition of the polymer was analyzed using 1H-NMR. The copolymerization composition DM and HEMA in the polymer was 10 molar percent and 90 molar percent, respectively. Average molecular weight (Mw) was $ 500000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material With a specific surface area of 1.47 m2/g, an average fiber diameter of 1.2 μm and a mass per unit area of 40 g/m2as the material. Covered the number was 18 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection, the level of hemolysis after filtration and removal efficiency of pathological prion protein by method 3 was tested in the ways described. The removal efficiency of leukocytes, the pressure during processing of blood, the level of hemolysis and the removal efficiency of pathological prion protein was 3,75, 15,0 kPa, 0.3 per cent and 0.5, respectively.

[Comparative Example 9]

Polymerization, purification, and drying was carried out similarly When the ERU 1, except the relationship of the charge of monomers 90 molar percent MMA and 10 molar percent DM. The copolymerization composition of the polymer was analyzed using 1H-NMR. Composition copolymerization of MMA and DM in the polymer was 90 molar percent and 10 molar percent, respectively. Average molecular weight (Mw) was $ 240000. The polymer solution at a concentration of 0.8 wt./vol.% in ethanol, used as a covering solvent, was coated polyester non-woven material With a specific surface area of 1.47 m2/g, an average fiber diameter of 1.2 μm and a mass per unit area of 40 g/m2as the material. Covered the number was 21 mg/m2per unit area on the entire surface area of the material. The removal efficiency of leukocytes, the pressure at the time of collection and the level of hemolysis after filtration was tested in the ways described. However, the test was discontinued because of the low fluidity of the blood product, a high pressure during treatment (more than 60 kPa) and fear of breakage of the hose and syringe, so that the measurement of the efficiency of removal of leukocytes and level of hemolysis was not possible.

[Comparative Example 10]

Conducted test 4 the efficiency of removal of pathological prion protein using a commercially available filter for removal of leucocytes to the product price is Inoi blood (WBF2, production Pall), and the removal efficiency of pathological prion protein amounted to 0.4.

Table 1 shows the results of Examples and Comparative Examples. In Table 1, the electron beam sterilization and sterilization by autoclaving denoted as EB and AC, respectively.

Applicability in industry.

Method of removing abnormal prion protein from a blood product according to the present invention is effective for preventing the transmission of blood for transfusion transmissible spongiform encephalopathy (TSE), caused by abnormal prion protein in the clinical field of blood transfusion, and to prevent adverse effects of blood transfusion caused by leukocytes.

1. Method of removing abnormal prion protein from a blood product, characterized in that it includes
filtering the product of blood through a filter filled with a carrier coated with the polymer formed by three elements, including 20 mol.% or more and 40 mol.% or less of the item originating from a hydrophobic polymerized monomer, 5 mol.% or more and 13 mol.% or less of the element originating from the polymerized monomer containing a basic nitrogen-containing part, and the element originating from the polymerized monomer containing a protonic neutral hydrophilic part as the balance; collecting filtered blood product.

2. Method of removing abnormal prion protein from a blood product according to claim 1, wherein the blood product is a product of whole blood, and the filter is subjected to sterilization by irradiation and then sterilized by autoclaving.

3. Method of removing abnormal prion protein from a blood product according to claim 2, characterized in that irradiation is a γ-rays or electron rays.

4. Method of removing abnormal prion protein from a blood product according to any one of claims 1 to 3, characterized in that the polymer is a vinyl polymer type.

5. Method of removing abnormal prion protein from a blood product according to claim 1, characterized in that the hydrophobic polymerized monomer, polymerized monomer containing a basic nitrogen-containing part, and the polymerized monomer containing a protonic neutral hydrophilic part, are derivatives of acrylic acid and/or derivatives of methacrylic acid.

6. Method of removing abnormal prion protein from a blood product according to claim 5, where the basic nitrogen-containing part is a tertiary amino group.

7. Method of removing abnormal prion protein from a blood product according to claim 5, where the proton neutral hydrophilic part is a hydroxyl gr the foam.

8. Method of removing abnormal prion protein from a blood product according to claim 5, where the basic nitrogen-containing part is a tertiary amino group, and the proton neutral hydrophilic part is a hydroxyl group.

9. Method of removing abnormal prion protein from a blood product according to claim 1 where the hydrophobic polymerized monomer, polymerized monomer containing a basic nitrogen-containing part, and the polymerized monomer containing a protonic neutral hydrophilic part, are derivatives of acrylic acid and/or derivatives of methacrylic acid, and filter is a filter for removing leukocytes.

10. Method of removing abnormal prion protein from a blood product according to claim 9, where the basic nitrogen-containing part is a tertiary amino group.

11. Method of removing abnormal prion protein from a blood product according to claim 9, where the proton neutral hydrophilic part is a hydroxyl group.

12. Method of removing abnormal prion protein from a blood product according to claim 9, where the basic nitrogen-containing part is a tertiary amino group, and the proton neutral hydrophilic part is a hydroxyl group.



 

Same patents:

FIELD: medicine.

SUBSTANCE: sampling of blood from patient's ulnar vein with first and second syringe-test-tubes is carried out. First syringe-test-tube is centrifuged with acceleration 250 g for 10 minutes. Formed plasma and coagulated blood in second syringe-test-tube are centrifuged with acceleration 1000 g for 10 minutes. Platelet-poor plasma is separated from platelet-enriched plasma. Activator of platelet-enriched plasma is prepared from supernatant fluid after centrifugation of coagulated blood and platelet-poor plasma, with weight ratio in fractions: 1:1, and 10% calcium chloride solution. The latter - in drop manner, mixing until required mixture concentration is obtained. Platelet-enriched plasma is mixed with activator with weight ratio in fractions 1:3 respectively.

EFFECT: application of method allows to realise more complete isolation of platelets from sampled blood, which increases efficiency, physiologicity and safety of obtained plasma.

2 cl

FIELD: medicine.

SUBSTANCE: system includes a kit of disposable elements for transportation of liquid, which are preliminarily connected or include aseptic connectors for creation of connections between them in aseptic way, or are adapted for being aseptically connected. Kit includes three sets of disposable sterile elements: set for collection, set for processing and set for transplantation, packed in blister packing on carrier, such as tray, which has one department, containing each connected with other set of said kit. System can be applied for obtaining platelet concentrate for separate application. Method of collection and processing of cell subpopulation includes collection of isolated cells in collection chamber, connected with isolation device, processing of cells in centrifugal operation chamber, which is the same chamber as the chamber for collection, or which is connected with collection chamber, and collection of processed cells in chamber for reinfusion, which also is the same chamber as the operation chamber, or which is connected with operation chamber, and delivery of processed cell subpopulations back to the patient.

EFFECT: application of invention makes it possible to ensure automatic processing in closed system in on-line mode.

17 cl, 9 dwg

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to surgery, and can be used in treating patients with secondary lymphedema of upper extremities. That is ensured by the introduction of autolymphocytes extracorporally processed with Roncoleukin and dissolved in normal saline in subcutaneous fat of an injured extremity in a projection of lymph node basins along a medial and lateral surface of forearm. The solution is introduced in 20 points by 0.5 ml containing 20 to 30 million cells per each point on the average. The procedures are thrice-repeated every 72 hours.

EFFECT: method allows to intensify lymphatic outflow from the injured extremity, to reduce paravasal inflammations of subcutaneous fat due to stimulating cell components of the immune system.

8 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: method involves the intravenous and/or intramuscular introduction of the drugs used in myelopathy, balneotherapy in the form of baths. Acupressure and manual therapy are applied in addition. The manual therapy requires such techniques, as ischemic compression, release phenomenon, fascial broach, postisometric relaxation of the muscles involved in a pathological process. Discontinuous therapeutic plasmapheresis is applied with using either Baxter, or Bekman, or Haemophoenix apparatuses, and/or extracorporal ultraviolet blood irradiations is ensured by the apparatus Isolda MD 73 M. The apparatus Cryo-Jet is used for cryotherapy with cooled dry air at the level of degenerative dystrophic changed vertebral motor cervical segments and muscle attachment points.

EFFECT: method improves clinical effectiveness due to fast pain management, enlarged range of active and passive motions, normalised muscular tonus, increased muscular strength and corrected vegetovascular disorders.

6 cl, 3 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, oncology, and can be used for treatment of malignant lung neoplasms in experiment. For this purpose performed is extracorporal irradiation of autoblood with red incoherent light λ=0.67 mcm dose W=3.06 J/cm2 in continuous mode with exposition duration to three minutes. After that 15-20 minutes later into said blood added is cyclophosphane in dose 40 mg/kg and the mixture is incubated for 40 minutes at T=37°C. After that it is re-infused into subclavian vein of the animal.

EFFECT: claimed method allows to reduce terms of anti-tumour treatment, increase efficiency of cyclophosphane action, reduce its toxic manifestations, increase animals' life term, increase non-specific anti-tumour resistance, activate anti stressor mechanisms.

2 tbl

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to gynecology and concerns the integrated treatment of tuboperitoneal infertility. That is ensured by low-intensity infrared lasing at power 150 Wt, wave length 0.89 mcm of the inguinal lymph nodes and a skin projection of the lumbosacral plexus. The exposure time is 1 min for each region for 1st session. Thereafter, the exposure time is increased to 3 minutes during 10 sessions. It is combined with local intraendometrial introduction of an antibacterial agent and an immunomodulator. Besides the treatment involves a number of plasmapheresis sessions and introduction of nonsteroidal anti-inflammatory drugs, antioxidants, desensitisers, and vaginal sanitation.

EFFECT: method provides effective restoration of menstruation and reproductive function.

2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to surgery and intensive therapy, and can be used in treating endotoxicoses caused by septic conditions. That is ensured by removal of a destruction area to be thereafter sanitised and drained. It is followed with lymphostimulating therapy by introduction of 0.25% Novocaine 60 ml with dissolved 0.1 g of Lydase and 1.0 ml of Pentoxifylline into the interspinous spaces in number of 1-3 injections. Further, discrete plasmapheresis in amount 400 ml for severe patients and 800 ml in moderate patients is applied in number of 1-8 procedures. The lymphostimulating therapy is alternated every second day with lymphotropic extracorporal pharmacoimmunotherapy with using a medicated mixture prepared by incubation of 40-60 ml of the patient's plasma with received plasma of the patient with Roncoleukin in dosage 500-1 million UN and Cefabol in dosage 1 g for 45 min at 37°C. In peritonitis and peritoneal sepsis, the medicated mixture is introduced in the bolus form in the retroperitoneal fat. The destructive processes in the pleural cavity require introduction of said mixture in the axillary cavity.

EFFECT: method provides endotoxin weakening in the bloodstream, intertissue spaces and cavities due to combined multidirectional local and system effect on the lymphoid system.

3 cl, 8 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to dermatology, and can be used for treating patients with psoriasis. Method includes drug therapy with application of hyposensibilising anti-histamine medications, hepatoprotectors, vitamins, local exfoliating and releasing ointments, general ultraviolet oirradiation. Method also includes plasmapheresis with division of blood into plasma and erythrocyte mass by centrifugation with removal of patient's plasma and erythrocyte mass reinfusion. Before returning erythrocyte mass into patient's organism it is processed with 200 ml of ozonised physiological solution with ozone concentration 2.5 mg/l. After that reinfusion of ozonised erythrocyte mass is carried out.

EFFECT: method allows to increase efficiency of psoriasis treatment due to reduction of PASI index level, correction of disturbed indices of hemostasis and lipid exchange.

2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to oncology, and can be used in contraction of anaemia accompanying the chemoradiation therapy. That is ensured by blood exfusion in amount 8-10% of the circulating blood volume, and the removed blood volume is replaced with crystalloids in the ratio 1:1. From the exfused blood, erythrocyte concentrate is recovered whereto Essentsiale N solution 5 ml is added; the prepared mixture is placed in a thermostat at temperature 37°C for 10 minutes. Then 100 ml of a physiologic saline is added to said mixture that is rocked. The prepared solution is introduced to the patient within 20 minutes with a simultaneous UV blood irradiation. Exfusion and extracorporal blood processing combined with UV irradiation are performed every second day, 2-3 sessions for the therapeutic course.

EFFECT: method allows reducing time for normalisation of the blood erythrocyte level and haemoglobin with maintaining the time intervals for chemoradiation therapy courses and without reducing the dosage of chemopreparations and radiation tumour load.

2 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly to gynaecology and urology, and concerns treating chronic recurrent urogenital bacterial and viral infections. That is ensured by producing lymphocyte self-cells of the patient's blood. Then said lymphocytes are cultivated with an immunomodulator. It is followed with intrauterine introduction of 0.5-2.0 ml of a lymphocyte suspension in an acceptable medium at their concentration 106-107 cel/ml.

EFFECT: without using antibacterial preparations, method provides evident antiviral and antibacterial effects, normalisation of the immune status with minimum by-effects.

5 cl, 4 ex, 3 tbl

FIELD: medicine.

SUBSTANCE: method of donor blood conservation includes mixing donor blood with hemoconservant glugicirum, additionally into composition of hemoconservant glugicirum included is 3-oxy-6-methyl-2-ethylpyridine succinate in amount 0.25 mg per 1 ml, separation of obtained conserves donor blood into plasma and erythrocyte mass, with further storage of erythrocyte mass.

EFFECT: invention allows to prolong storage terms of morphofunctionally full value erythrocytes in donor erythrocyte mass up to 30 days.

2 dwg, 10 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to anesthesiology and resuscitation, and can be used in states accompanied by massive blood loss. For this purpose, 5 and 3 days prior to a surgery, autoblood is sampled in amount 10 % of circulating blood volume (CBV) in each sampling to be divided on plasma and erythrocyte concentrate. For 5 preoperative days, iron preparations are administered in a therapeutic dose. Also, the surgery is preceded with analysing patient's fluid deficiency. 40 minutes prior to the surgery, 12.5 % dicynone 500 mg, one volume of autoplasma are introduced, and infusion of a rated dose of 5 % glucose and 6 % hydroxyethyl starch (HES) 500 ml is started. If the intraoperative blood loss is suggested to be 15-30 % of the CBV, additionally 6 % HES 250 ml, prednisolone in dosage 2-4 mg/kg of body weight and one volume of erythrocyte concentrate are administered. If the estimated intraoperative blood loss exceeds 30 % of the CBV, the second volumes of autoplasma and erythrocyte concentrate, another introduction of dicynone in the same dosage, 6 % HES 250 ml, prednisolone in dosage 7 - 10 mg/kg of body weight are required. Within 30 and 60 minutes after the surgery, dicynone is injected in the same dosage. 5 hours after the surgery, filter drainage fluid is returned. 6 hours after the surgery, coagulation time is determined, and if observing no hypocoagulation, Clexane is introduced in a preventive dose.

EFFECT: method allows to provide early activation of erythropoiesis combined with improved erythrocyte morphology and blood haemostatic function, considerably reduced risk of complications connected with massive transfusion of donor blood products, as well as prevented edema of interstitial spaces and development of multiple-organ-failure syndrome due to maintained effective transcapillary exchange.

5 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to surgery, and can be used in treating patients with secondary lymphedema of upper extremities. That is ensured by the introduction of autolymphocytes extracorporally processed with Roncoleukin and dissolved in normal saline in subcutaneous fat of an injured extremity in a projection of lymph node basins along a medial and lateral surface of forearm. The solution is introduced in 20 points by 0.5 ml containing 20 to 30 million cells per each point on the average. The procedures are thrice-repeated every 72 hours.

EFFECT: method allows to intensify lymphatic outflow from the injured extremity, to reduce paravasal inflammations of subcutaneous fat due to stimulating cell components of the immune system.

8 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: invention relates to therapeutic-preventive medication for oral cavity hygiene and is used in dentistry. Therapeutic-preventive medication for oral cavity hygiene, which represents mixture of dry pantohematogen and dry extract of bergenia taken in specified ratio.

EFFECT: medication possesses efficient antibacterial, immunomodulating, anti-inflammatory, keratoplastic action, does not contain preservatives and dyes.

2 dwg, 3 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to anaesthesiology, and can be used as an anaesthesia care for surgeries, including oncotomy of various localisations. That is ensured by preoperative blood sampling in the amount 15 ml in a syringe with Solutio Glugicirum. Thereafter, dalargin 0.85 mcg/kg diluted in the same preservative is added in the syringe, and the prepared mixture is incubated at temperature 37°C for 30-35 minutes. Immediately before the induction, an anaesthetic is added with 1/3 dose of dalargin with autoblood and verapamil in dosage 0.9 mcg/kg. It is followed with the anaesthetic induction with thiopental sodium, ketamine, dormicum. Before the traumatic stage, 1/3 dose of dalargin with autoblood and verapamil in dosage 0.9 mcg/kg are introduced. The most traumatic stages require thiopental sodium, propofol, dormicum, phentanyl to be introduced. 30-40 minutes prior to the surgery completed, 1/3 dose of dalargin with autoblood are introduced in combination with actovegin in dosage 5.6 mg/kg.

EFFECT: technique enables adequate analgesia during the whole intra- and postoperative period combined with reduced dosage of the administered narcotic analgesics due to the optimised circulatory system, activated stress limit processes and normalised neurovegetative body functions.

2 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to otolaryngology, and deals with treatment of chronic recurring inflammatory diseases of nasal mucosa and paranasal sinuses. Method includes preliminarily obtaining of autologic lymphocytes from venous blood of patient, cultivation of obtained lymphocytes together with immunomodulator, preferably immunofan. After that from 0.1 to 3.0 ml of lymphocyte suspension, which contains 106-107 cells/ml, is introduced into paranasal sinuses with intervals between introductions from 2 to 7 days.

EFFECT: method is efficient, including cases of recurring polyps rhynosinutisis, allows to reduce treatment terms, increase remission terms, reduce frequency of appearance of symptoms of main and accompanying disease with reduction of need in other therapy.

4 cl, 3 ex, 6 tbl

FIELD: medicine.

SUBSTANCE: invention relates to medicine, ophthalmology, and can be used for treatment of optic atrophy in children of age from 1 to 6 months. For this purpose regions of large and small fontanelles of head and upper sympathetic ganglia at the level of cervical spine at C1-C2 level are exposed to polarised light of apparatus "Bioptron". Exposures to light are carried out daily during 10 days in therapeutic dose 12 J/cm2 from the distance 5 cm from skin surface with light spot diametre 5 cm, irradiation intensity 40 mW/cm2 exposure duration 30 seconds on region of fontanelles and 1 minute on regions of ganglia. On finishing light impact nootropic medications cortexin and/or actovegin or cerebrolisin are introduced in age dose. Courses of treatment are carried out from 1 month to 6 month age until desired therapeutic effect is obtained.

EFFECT: method allows to improve cerebral and intraocular blood supply and metabolic processes of brain and optic nerve, improve delivery of nootropic medications to optic analyser, obtain therapeutic concentrations of medications without side effects.

1 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to oncology, and can be used for treating brain glioma. For this purpose after tumour ablation glucocortocoids are introduced intramuscularly daily and radiotherapy is carried out. Before beginning irradiation and in process of treatment every 7-10 days blood serum is applied on transparent surface, dried at 20 - 25°C and relative humidity 55 - 60% and microscopic image of blood serum fascies is analysed. If in sample in process of irradiation leaf-shaped structures appear or their number increases in comparison with sample before irradiation less than 2 times, additionally trental is introduced intravenously in dose 100-200 mg twice per week. If number of said structures in sample increases 2 times and more trental is introduced intravenously in dose 200-300 mg twice per week and actovegin is introduced in dose 400 mg daily. Medications are introduced before finishing course of radiotherapy.

EFFECT: increase of treatment efficiency and possibility to carry out complete course of radiotherapy due to individual control of state of microcirculatory brain bed and depth of its injuries, cause by radiotherapy, which allows to carry out their drug correction in due time in accordance with individual programme.

4 dwg, 2 ex

FIELD: medicine.

SUBSTANCE: invention concerns microbiology and biotechnology. There is offered a method for prepared activated mononuclear leukocytes involving the use of interleukin-2 in combination with cyclodextrin for mononuclear cell activation, a lymphocyte and macrophage concentrate recovered from donor or patient blood is activated in vitro with a complex containing a pharmacopeia preparation of recombinant interleukin-2 within the range of suboptimal doses 50-100 MU in 1 ml of the culture medium and β-cyclodextrin in concentration 1×10-5-1×10-3 M in the culture medium at the molar ratio of components either 1:1, or 1:3, or 1:10, the lymphocyte and macrophage concentrate of donor or patient blood is processed in vitro with a complex containing a pharmacopeia preparation of recombinant interleukin-2 within the range of suboptimal doses 50-100 MU in 1 ml of the culture medium and β-cyclodextrin in concentration 1×10-5-1×10-3 M in the culture medium the molar ratio of components either 1:1, or 1:3, or 1:10.

EFFECT: invention provides the preparation of IL-2/CD-MNL with high cytotoxic activity to tumour cell lines; 10-fold reduction of IL-2 doses with preserved clinical effectiveness of the activated cells allows reducing the intensity of systemic by-effects of the preparation, and also the use of immunotherapy in debilitated oncological patients, including in the patients with a hyperreactive response to the IL-2 introduction.

2 ex, 1 tbl, 10 dwg

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to methods of obtaining deposited lymphokine-activated killer cells (LAKC) for treatment of oncologic diseases. Claimed is method, according to which mononuclear lymphocytes (MNL) are isolated from peripheral blood of malignant effusion by centrifugation on one-step gradient of ficoll with density 1.06-1.08 g/cm3, LAKC are generated and concentrated by successive precipitation of MNL by centrifugation, re-slurring it in RPMI 1640 or DMEM medium with addition of 2-10% AB human serum, interleikin-2 in concentration 0.5-1.0 mlnIU/ml and incubation in CO2 incubator for 48-72 hours, after that suspension of obtained LAKC is deposited in culture medium in amount 2 mln LAKC per 200 ml of medium on sterilised and washed with culture medium porous titanium carriers with 55-60% porosity.

EFFECT: invention allows to increase efficiency of obtained medication for local and local-regional immunotherapy of oncologic patients.

1 ex, 8 dwg, 2 tbl

FIELD: medicine, surgery.

SUBSTANCE: at the background of basic therapy in complex therapy of acute pancreatitis one should introduce ceruloplasmin to be applied at the dosage of 600-1000 mg/d for 5 d. If necessary, the course of ceruloplasmin introduction should be repeated. This method provides pancreatic tissues viability in case of pancreonecrosis by increasing efficiency of correction the endogenous intoxication and decreasing the number of complications in the course of therapy conducted.

EFFECT: higher efficiency of pharmacological correction.

3 ex

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