Method of obtaining fibrinogen concentrate
SUBSTANCE: invention relates to biotechnology of obtaining haemostatic medications. Claimed is method of separating purified fibrinogen concentrate, free of viruses and ballast proteins. Solubilisation of cryoprecipitate of fresh frozen human plasma is realised. Fibrinogen is precipitated with 20-30% PEG solution. Separated sediment is dissolved in buffer with sodium citrate and sodium chloride. Virus inactivation of solution by solvent-detergent method is carried out in presence of 1-3% Tween-80 and 0.1-1.5% of tri-n-butylphoshate. Obtained concentrate is purified from products of virus inactivation and solvent-detergents by triple extraction with liquid paraffin. After that, obtained fibrinogen concentrate is re-precipitated with 1.0-2.5 M glycine solution. Sterile filtration and lyophilic drying with further corking of lyophilisate under vacuum and thermal inactivation are performed.
EFFECT: invention makes it possible to obtained lyophilised form of fibrinogen concentrate with approximately 55% output.
The invention relates to pharmaceutical industry, namely to the biotechnology production of hemostatic drugs. Fibrinogen is synthesized in the liver and secreted in plasma in concentrations sufficient for coagulation. Fibrinogen is a soluble glycoprotein that is converted under the action of thrombin in the fibrin network of fibres which is the basis of the bunch, providing hemostasis. Besides the hemostatic function of fibrinogen is involved in wound healing, tumor development and metastasis (Handbook for medical laboratory technicians in research methods of plasma hemostasis. The blood coagulation factors. 2008, Moscow, p. 7).
Congenital deficiency of fibrinogen (hypofibrinogenemia) can disrupt homeostasis and lead to bleeding. More common is the acquired deficiency of fibrinogen, which may be a consequence of hemodilution, blood loss, disseminated intravascular coagulation (DIC) and sepsis (Bratchikov A. M. Clinical problems of fibrinolysis. 1993, Kiev, pp. 151-172).
Hereditary or acquired fibrinogen deficiency causes the development of hypofibrinogenaemia, the main method of treatment and prevention which is replacement therapy with preparations of fibrinogen" (Zubairov D. M. Fibrinogen and fibrin. In: Molek�regular basis of blood coagulation and clot formation. 2000, Kazan, pp. 1-32).
Substitution therapy in patients with hypofibrinogenemia and its prevention using preparations containing fibrinogen.
In addition to replacement therapy drugs based on the received fibrinogen concentrate can be used in surgical practice as a component of fibrin glue. The composition of this glue allows you to perform operations on large wound surfaces, whether the operation in areas with a high degree of vascularity, or processing of large surfaces burns that accelerates the regeneration of epithelial tissues (Zilbert A. P. Blood loss and blood transfusion. Principles and methods of bloodless surgery. 1999, Petrozavodsk, pp. 55-62).
To the problems of production of drugs on the basis of fibrinogen from human blood plasma include:
- the possibility of infecting the recipient with hepatitis viruses (A, B, C), human immunodeficiency, parvovirus, syphilis and other pathogens,
- the possibility of transfer to the recipient with a certain group of blood group present in the preparations (concentrates average purity) of antibodies to antigens of red blood cells from other blood groups,
- contamination of drugs on the basis of fibrinogen ballast proteins,
To solve these problems develop appropriate technological approaches. Thus, the preparation of the pool of plasma is carried out OTB�R carefully screened healthy donors, which determine possible infection with hepatitis viruses, human immunodeficiency, etc. However, to fully guarantee the absence of viral infection in the process of obtaining concentrates of fibrinogen carry out viral inactivation.
A method of producing a concentrate of fibrinogen by salting out using alcohol-salt cold deposition" (blood products. Instructional and methodological materials for the control and production. - M., 1976, pp. 145-202). This method requires a large time and energy costs, preparation of complex buffer salt solutions and the need for their subsequent laundering, making the process cumbersome, and the duration of the procedure leads to a significant loss of product during receipt. Method can not be recommended for the production of a preparation from small amounts of blood.
Deposition of fibrinogen ammonium sulphate also requires subsequent purification of the product from salt solutions, which leads to additional technological difficulties" (Patent RU №2062103). Concentrates of fibrinogen obtained by cryoprecipitate, successfully applied in foreign surgical practice" (Patent RU №2062103), however, achieved the concentration of fibrinogen remains relatively low. The closest to the claimed method is a method of producing drug concentrations�ATA fibrinogen, presented in the Patent US 4960757.
The method consists in obtaining a concentrate of fibrinogen precipitate containing fibrinogen, obtained in the process for preparation of factor VIII is carried out according to the patent German Patent No. 2916711. The feedstock is dissolved in a buffer containing NaCl, by heating to 37°C and stirring to maintain pH=7,5 2N NaOH. Further to the fibrinogen concentrate was added a solution of CaCl2.2H2O and sucrose to a concentration of 60% in the final solution. Then, the solution of fibrinogen with stabilizers subjected to thermal pasteurization in a water bath at 60°C, to inactivate viruses and other pathogens. After pasteurization, the fibrinogen solution is diluted with buffer containing sodium chloride and sodium citrate and dwukrotnie periostat glycine. The disadvantage of this method is to reduce the concentration of fibrinogen in the final product of the partial polymerization at the stage of thermal inactivation. Formed at this stage, the fibrin is separated double-glycine precipitation. Thus the yield of fibrinogen in the final stage becomes smaller.
The aim of the present invention to provide a method for producing a purified fibrinogen. This goal was achieved by the development of a technology for providing a lyophilized form of a concentrate FiBr�of novena. The advantage of this method of allocation is the use of a buffer solution containing anticoagulate agent is sodium citrate. Just one advantage is the use of Tween-80 (monooleate polyoxyethylenesorbitan) and TnBP (tri-n-butylphosphate) as detergents when conducting viral inactivation. In this method also be in the process of purification of fibrinogen concentrate from the products of the process of viral inactivation.
The technology provides for the dissolution of the feedstock (cryoprecipitate fresh frozen human plasma) in water in the presence of 1-3 units of heparin, treatment of a solution of cryoprecipitate gel of aluminium hydroxide, the precipitation of fibrinogen 20-30% solution Page, the subsequent dissolution of selected sediment in the buffer with sodium citrate and sodium chloride, viral inactivation solution solvent/detergent Tween 80 and TnBP within 6-10 hours, removal of products of inactivation and inactivating agents extraction solution of liquid paraffin, purified from ballast two proteins by precipitation with glycine concentration of 1-2.5 mol/l, subsequent sterile filtration, filling and lyophilization.
The development of separation technology of fibrinogen first conducted in the laboratory with subsequent optimization of the chosen method, and then scaled textures in the conditions experienced PR�production. The production of the preparation of fibrinogen was carried out in accordance with the rules of good Manufacturing Practice (GMP), standard operating procedures (SOP) and necessary conditions of sterility".
Raw material for production of fibrinogen concentrate was frozen cryoprecipitate fresh frozen human plasma.
At all stages of production raw materials, intermediate products and targeted drug tested on the content of fibrinogen by the method of Claus.
Below is a specific example of the invention.
1. The feedstock (cryoprecipitate fresh frozen plasma) in an amount of 100 g was dissolved in three times the volume of distilled water containing 1-3 units of heparin and add 3% gel Al(Oh)3(ratio: 1 kg suspended cryoprecipitate to 10 g of aluminum hydroxide).
2. Next, to this mixture was added 30% solution of Page, to a concentration of 3%, and the pH of the solution was adjusted to a value close to the isoelectric point of fibrinogen (6,6-6,8) acetic acid. Under the action of Paga fibrinogen precipitate, which was separated from the supernatant by centrifugation. Next, the supernatant is transferred to the production of purified factor VIII.
3. The precipitate containing fibrinogen was dissolved with vigorous stirring in a tenfold volume of buffer containing 10 mmol sodium citrate and 150 mmol of sodium chloride, maintaining the pH at 7.8 to 8.0 with 0.5 M solution of Tris-buffer, and a temperature of 30-35°C. the solution was Then centrifuged for clarification at 4000 rpm for 10 minutes.
4. Viral inactivation of fibrinogen carried out in the presence of 1% Tween-80®and 0.3% TnBP, incubating the resulting solution at room temperature with continuous stirring for 6-10 hours.
5. Removal of products of inactivation and inactivity agents is carried out by extraction of fibrinogen solution paraffin oil. For this to virusinaktivierung fibrinogen concentrate is added mineral oil in the ratio of 2:1 and intensively stirred for 20 min. the mixture was centrifuged at 2000 rpm for 10 minutes and the aqueous layer is separated. This procedure is carried out three times.
6. The separation of fibrinogen from the ballast proteins is carried out by precipitation of the obtained concentrate of fibrinogen glycine, bringing its concentration in the solution to 1.5 mol/L. thus, a precipitate of fibrinogen, and most of the ballast proteins remain in solution. Of postural fibrinogen was separated by centrifugation at 4000 rpm for 10 minutes and dissolved in a solution containing 10 mmol sodium citrate and 0.15 mol of sodium chloride. The procedure was repeated twice, each time washing the precipitate fibrinogen cold 1.5 M solution of glycine.
After the resultant deposition of fibrinogen dissolved in the buffer to the observed concentration�radio 25 g/l, sterile-filtered, poured into bottles and lyophilizers. The obtained lyophilisate sealed under vacuum, vials crimped aluminum caps and subjected to thermal inactivation at 80°C for 72 hours.
The yield of fibrinogen is about 55%.
A method of separating a purified concentrate of fibrinogen, free from viruses, namely, to solubilize the starting material, represented by cryoprecipitate fresh frozen human plasma, deposition of fibrinogen 20-30% solution of PEG, selected by dissolving the precipitate in buffer with sodium citrate and sodium chloride, virus inactivation of a solution solvent-detergent method in the presence of 1-3% Tween-80 and 0.1 to 1.5% tri-n-butylphosphate, purifying the resulting concentrate products from viral inactivation and solvent-detergent extraction liquid paraffin conducted three times, subsequent precipitation of the obtained concentrate of fibrinogen 1,0-2,5 M solution of glycine, sterile filtration and freeze-drying with subsequent capping of the extract under vacuum and terminatively.
SUBSTANCE: invention relates to the field of biochemistry, in particular to recombinant factor VIII, which contains one or more mutations, resulting in an increased stability of both the factor VIII and factor VIIIa, as well as to a pharmaceutical composition for treating haemophilia containing it. Also described is a molecule of nucleic acid, coding the said recombinant factor VIII, and an expression vector and host-cells, containing the said molecule of nucleic acid. The invention also relates to a method of obtaining the said factor VIII, as well as to its application in the method of treating haemophilia A in an animal.
EFFECT: invention makes it possible to obtain a biologically active factor VIII with an increased stability.
50 cl, 12 dwg, 5 tbl, 9 ex
SUBSTANCE: method of purifying coagulation factor VIII protein from a solution involves: a) contacting the protein with a multimodal resin or a mixed action-type resin containing ligands having a hydrophobic portion and a negatively charged portion; b) elution of the protein with an elution buffer containing at least 1.5M salt and at least 40% (wt/vol) ethylene glycol, propylene glycol or mixture thereof, and calcium ions. The method is a single-step chromatographic process where the coagulation factor VIII protein is captured and does not require adjusting pH or electroconductivity at the feeding step. The method of stabilising the coagulation factor VIII protein, where the coagulation factor VIII protein is captured and stabilised when the protein passes once through a column with a multimodal resin containing ligands having a hydrophobic portion and a negatively charged portion, and elution of the protein with an elution buffer containing at least 1.5M salt and at least 40% (wt/vol) ethylene glycol, propylene glycol or mixture thereof, and calcium ions.
EFFECT: invention reduces the volume of the column about 250-fold and a coefficient of purification equal to 30, high stability of the protein product.
2 cl, 4 dwg, 4 tbl, 7 ex
SUBSTANCE: in order to reduce content of dimers in a composition which contains a factor VII polypeptide or a version of factor VII polypeptide, pH of said composition is brought to a value ranging from 5 to 10.0; the composition is heated to temperature ranging from 20°C to 50°C for 10 minutes to 72 hours, followed by cooling to temperature not higher than 5°C.
EFFECT: low dimer content in a factor VII polypeptide composition by heat treatment.
8 cl, 4 dwg, 3 ex
SUBSTANCE: method to produce polypeptide of factor VIII provides for cultivation of cells of mammals, which express polypeptide of factor VIII, under conditions, suitable for expression of the specified popypeptide of factor VIII, which include a cell cultural medium with O-phospho-L-serine (OPLS) and extraction of the expressed polypeptide of factor VIII from cells of mammals with the help of suitable means.
EFFECT: invention makes it possible to produce popypeptide of factor VIII with increased specific activity.
11 cl, 3 dwg, 4 tbl, 2 ex
SUBSTANCE: invention relates to biotechnology and can be used to extract plasminogen or plasmin. A mixture containing fibrinogen and plasminogen or plasmin is deposited onto a chromatographic column containing an insoluble affinity chromatography matrix which is covalently cross-linked with tranexamic acid. Plasminogen or plasmin is then eluted from the affinity chromatography matrix with an aqueous solution containing a sufficient amount of a ligand which competes with plasmin or plasminogen for binding sites of said matrix which is bonded with tranexamic acid. Tranexamic acid is covalently bonded with said matrix through a linker which lies between the matrix and tranexamic acid and whose length is more than three carbon atoms.
EFFECT: invention increases efficiency of extracting plasminogen or plasmin from a mixture in the presence of fibrinogen.
10 cl, 1 dwg, 13 tbl, 6 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention concerns area of molecular biology and biochemistry, and can be used in medicine. There is offered mutein conjugate of the blood coagulation factor VIII (FVIII) wherein a residue not being cysteine in position 41, 129, 377, 388, 468, 491, 556, 1804, 1808, 1810, 1812, 1813, 1815 and/or 2118 is substituted by a cysteine residue with polyethylene glycol (PEG) where a PEG molecule is bound with a polypeptide in a mutant cysteine residue.
EFFECT: improved pharmacokinetic properties of the FVIII as an ingredient of the conjugate under the invention with preserved a procoagulant factor activity allows presenting new FVIII PEG-muteins for producing of a pharmaceutical compositions for treating hemophilia.
12 cl, 38 dwg, 8 tbl, 1 ex
FIELD: chemistry; biochemistry.
SUBSTANCE: invention relates to biotechnology and specifically to obtaining versions of glycoprotein IV alpha polypeptide of human thrombocytes (GPIbalpha) and can be used in medicine to treat vascular disorders. Using a recombinant technique, a polypeptide is obtained, which contains substitutes in SEQ ID NO:2 selected from: Y276F K237V C65S; K237V C65S; Y276F C65S; or Y276F Y278F Y279F K237V C65S. The obtained polypeptide is used to inhibit bonding of leucocytes to biological tissue or for treating disorders associated with activation of thrombocytes.
EFFECT: invention enables to obtain GPIbalpha polypeptide which bonds with von Willebrand factor with affinity which is at least 10 times higher than in natural GPIbα polypeptide, and also has low affinity for bonding with alpha-thrombin, lower aggregation and/or high resistance to proteolysis relative the polypeptide with SEQ ID NO:2.
41 cl, 3 dwg, 8 ex
FIELD: chemistry, biochemistry.
SUBSTANCE: invention refers to biochemistry and can be used in pharmaceutical industry for mixture purification of natural mixtures containing plasminogen and fibrinogen from plasminogen. Plasminogen is removed from mixture containing plasminogen and fibrinogen using insoluble chromatography matrix that is covalently cross-linked with tranexamic acid through amino group with linker of length exceeding three carbon atoms. Therefore mixture mentioned above is applied on chromatographic column containing specified insoluble matrix. Then column is washed with neutral solution containing salts, while unbound material is collected.
EFFECT: extended technical feasibilities of purification of natural mixtures containing plasminogen and fibrinogen from plasminogen thus keeping original amount of fibrinogen in mixture.
15 cl, 1 dwg, 13 tbl, 6 ex
FIELD: biotechnology, in particular production of modified swine factor VIII (POL1212).
SUBSTANCE: DNA molecule encoding of modified swine factor VIII is cloned in expression vector, having functionality in mammalian cells. Modified swine factor VIII protein is obtained by cultivation of mammalian cell line BHK CRL-1632 (ATCC), BHK 1632, or CHO-K1, transfected with vector. Therapeutic composition for treatment of subjects suffering from deficit of factor VIII, such as haemophilia, contains effective amount of swine factor VIII protein.
EFFECT: effective agent for treatment of factor VIII deficit.
13 cl, 8 dwg, 7 ex
SUBSTANCE: method comprises the steps of destroying the bodies of inclusion, renaturation and purification of protein. Before renaturation, the preliminary purification of protein is carried out using chromatography on Q-Sepharose and SP-Sepharose using the combined columns with Q- and SP-Sepharose. After renaturation chelate and ion-exchange chromatography of recombinant prourokinase M5 is carried out without intermediate elution of the target protein with use of metal-chelate sorbent activated with ions Co2+ or Zn2+.
EFFECT: invention enables to select prourokinase M5 from inclusion bodies, comprising the present prourokinase, with high yield.
4 cl, 2 tbl, 4 ex
SUBSTANCE: invention relates to the field of obtaining and separation of single-domain molecules (SDAB). Described is a method of the separation or purification of the SDAB molecule, which represents a trivalent molecule of a ATN-103 nanobody, targeting TNFα and HAS, from a mixture, containing the said SDAB molecule and one or more polluting substances. The mixture is brought in contact with a cation-exchange carrier under conditions, which make it possible for the SDAB molecule to bind with the carrier or be absorbed on the carrier. One or more polluting substances are removed and SDAB is selectively eluted from the carrier. The conductivity of a conditioning medium (CM), used for the carrier loading, constitutes from approximately 12 to 9 mS/cm and pH under conditions of loading is corrected to a value from 4.0 to 4.3. The buffer for elution corresponds to approximately 50 mM of sodium chloride or less and has pH from approximately 5.5 to 7.2. Disclosed is a method or a process of obtaining recombinant SDAB of ATN-103. A host-cell is supported in the conditions at which recombinant ATN-103 SDAB is expressed. The mixture of molecule SDAB and one or more polluting substances is obtained. ATN-103 SDAB is purified or separated with the application of cation-exchange chromatography, as said above.
EFFECT: application of the invention provides new methods of the separation or purification of the nanobody, which can be applied in obtaining the ATN-103 nanobody.
19 cl, 4 dwg, 6 ex
SUBSTANCE: method of obtaining of a complex of antimicrobic peptides of an insect includes infecting of adipose body of an insect at a larval instar with Micrococcus luteus A270 and Escherichia coli D31 bacteria with the subsequent extraction of adipose body of an insect at a larval instar. The adipose body of an insect is placed into a nutrient medium containing water solution of sugars, inorganic salts and the antibiotic meropenem in pre-set ratio and incubated during a day with the subsequent elution of the complex of antimicrobic peptides of an insect from cultural liquid by the method of reverse-phase chromatography on the column Vydac C18 at the linear gradient of acetonitrile from 0% up to 50%.
EFFECT: invention allows to simplify a method of obtaining antimicrobic peptides.
5 dwg, 4 ex
SUBSTANCE: invention refers to peptide chemistry and concerns producing tripeptide diacetate H-β-Ala-Pro-DabNHBzl referred to a biologically active compound used in cosmetic industry as an active component for cosmetic products, particularly for stimulating skin rejuvenation, tightening and prevention. The method is based on the 6-staged synthesis and is free from the stages of setting and releasing the protective groups. The method involves proline β-chlorpionyl chloride acylation followed by producing N-(3-chlorpropionyl)proline pentafluorphenyl ester in the presence of N,N'-dicyclohexyl carbodiimide. The above pentafluorphenyl ester is condensed thereafter with glumatic acid monomethyl ester to produce N-(β-chlorpropionyl)-Pro-Glu(δ-OMe)OH. That is followed by benzylamine amidation in a combination with ammonolysis and chlorine substitution by an amino group. That enables producing the tripeptide β-Ala-Pro-Glu(δ-NH2)NHBzl; Hofmann rearrangement is conducted with the use of iodo-benzene diacetate to produce a target product.
EFFECT: method is characterised by simplicity, effectiveness; it is cost-effective and uses more accessible and cheap agents.
SUBSTANCE: method of obtaining SSI comprises the following steps. The strain Yersinis pestis KM 1279 is grown on 1.5% agar LB, the bacteria are washed three times with cold buffered normal saline. The bacteria are pelleted by centrifugation, suspended in a solution of 5 mM NaOH, kept at 37°C for two hours and the cells are pelleted by centrifugation. The supernatant is selected and the procedure as repeated three times, three supernatants are combined and filtered through the nitrocellulose membrane. The filtrate is extracted three times with the mixture of chloroform-methanol-water in a ratio of 5:2:1. The chloroform fractions are separated by centrifugation, combined and freed from water-soluble impurities. The aqueous fraction is separated by centrifugation and removed, and the chloroform fraction is dried in a vacuum rotary evaporator and the dry preparation SSI is obtained. The proposed SSI is characterised with brown colouring of dry crystals, hydrophobic properties, fluorescence in ultraviolet, lipopeptide nature, the presence of iron ions, the molecular weight of 380.6 Da.
EFFECT: inventions enable to obtain the natural regulator of virulence of plague agent.
2 cl, 6 dwg, 6 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology. What is presented is a method for preparing a recombinant protein of type III interferon-like factor (ILF III) of the producing strain E. coli. The inclusion bodies E. coli are washed and dissolved with using 2% aqueous γ-cyclodextrin. That is followed by the sequential Ni-Sepharose, Q-Sepharose and SP-Sepharose chromatographic procedures. Refolding of a target protein is performed with using a mixture of cysteamine and cystamine at pH 10.5. The Amberchrome Profile XT20, Amberchrome Profile HPR10 and Kromasil 300-5C18 chromatographic procedures are sequentially performed.
EFFECT: invention enables optimising the ILF III purification environment at the stage of washing and dissolving the inclusion bodies Ecoli and provides 12% target protein yield.
3 dwg, 4 ex
SUBSTANCE: invention relates to biotechnology, specifically to immunostimulating compounds and may be used in medicine. An immunostimulating peptide of an amino acid sequence XLYDKGYTSKEQKDCVGI, where N-terminal X is N-acetylalanine, and may be covalently linked to fatty acids, selected from C2-C25, to form PDAG (peptidyl-2,3-diacylglycerides). The resulted compound may be included in pharmaceutical formulations for stimulating an immune response.
EFFECT: invention provides efficient stimulation of an immune response in subjects and may enhance the immunogenicity of the antigenic peptide when administered with PDAG.
29 cl, 10 dwg, 9 ex
SUBSTANCE: method of production of peptides is proposed. Yeast autolysis is carried out. The cell membranes are separated by centrifugation. The autolysate is purified on the gel sulphocationite in the hydrogen form, containing 12-16% divinylbenzene. The resulting peptide aqueous solution is passed sequentially through the gel anion-exchange material to obtain the solution at pH 2.0-2.6, and then through the gel cation-exchange material with the divinylbenzene content of 1-2% or macroporous cation-exchange material.
EFFECT: obtaining highly purified peptides that have biological activity.
SUBSTANCE: invention relates to purification of various gamma-carboxylated olypeptide forms with application of ion-exchange chromatography. In particular, in accordance with invention, claimed is method of purification of polypeptide, which has desirable content of gamma-carboxyglutaminic acid, from sample, containing mixture of said polypeptide versions, which have different content of gamma-carboxyglutaminic acid, with the claimed method including stages: (a) loading said sample on anion-exchange chromatographic material; (b) elution of said polypeptide with application of solution with pH lower than 9.0, containing at least one salt, selected from ammonium acetate, ammonium chloride and sodium acetate; and (c) selection of fraction, obtained after said elution, with polypeptides in said fraction having desired content of gamma-carboxyglutaminic acids.
EFFECT: claimed is method of purifying polypeptide, which has desirable content of carboxyglutaminic acid.
8 cl, 13 dwg, 6 tbl, 7 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology. What is presented is a composition of an antibody for treating HER2-positive cancer containing an active antibody presented by an antibody characterised by the fact that it has a variable light and heavy chain domain, and its acidic variants, namely: glycosilated, deaminated variants, as well as a variant with a reduced disulphide bond, a syalylated variant and a irreducible variant. A number of acidic variants makes less than approximately 25%. There are described a pharmaceutical composition containing this composition, for treating HER2-positive cancer and a method for preparing the composition involving the evaluation of the acidic variants and verification of the fact that their number makes at least than approximately 25%.
EFFECT: using the invention provides the new composition, wherein the antibody and its acidic variants have the pharmacokinetic parameters that can find application in treating HER2-positive cancer.
14 cl, 17 dwg, 1 tbl, 1 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions refers to medicine and concerns a method for making an activated leukocyte composition involving human leukocyte incubation, exposure to hypotonic shock and addition of physiologically acceptable saline in an amount sufficient for the reconstitution of isotonicity, to the leukocytes. The group of inventions also concerns using: the activated leukocyte composition in preparing a drug for wound healing; a wound healing dressing containing the above composition.
EFFECT: group of inventions provides preparing the composition containing a 90 times increase of the leukocyte count as compared to the known state-of-the-art methods.
16 cl, 3 dwg, 6 tbl, 3 ex