The method of obtaining the immunoglobulin preparation
(57) Abstract:The invention relates to medicine, in particular to immunology. The method of obtaining the immunoglobulin preparation is salt extraction alcohol precipitate of human serum, clearing the lipoproteins, followed by separation of the target product, adding a stabilizer, containing glucose, and sterilizing filtration, with a selection of immunoglobulins conduct alcohol precipitation, as stabilizer, a mixture of glycine and glucose at a ratio (1-1,5):1, a solution of immunoglobulins treated sorbent glass, at the stage of selection of the target product are conducting a flow-zonal centrifugation, and before sterilizing pass filtering the solution through a porous glass filter. Technical result: increasing output and lowering the cost of the drug. The invention relates to medicine, in particular to immunology, and can be used for immunotherapy and prophylaxis of bacterial and viral diseases.A method of obtaining immunoglobulin drug protected by a patent SU 1767742, class a 61 K 37/04, published 11.06.90.Way to bookmark and purification of immunoglobulins by dissolving the precipitate in 0.05 M acetate buffer, pH 4.3 and treatment kaprilat sodium to a final concentration of 0.5%, pH of 4.8, the deposition of immunoglobulins alcohol, adding 0.15 M glycine and 2% sucrose, holding sterilizing filtration and the addition of pectin at a concentration of 0.3-0.5 g per 1 g of immunoglobulin.The disadvantage of this method is the presence of impurities kaprilat sodium in the final product and the instability of the resulting solution immunoglobulin able to be kept after sterile filtration prior to bottling no more than 10 hoursClosest to the claimed technical essence and the achieved result is a method for immunoglobulin preparation (TRC) from sludge B (III fraction Kona) (patent RU 2060034, class a 61 K 39/395, published 06.09.91)
The method comprises the salt extraction of the alcohol precipitate B serum (III fraction Kona) in 0.9% sodium chloride solution. Clearing of lipoprotein extract sediment B are chloroform (10% by volume), followed by deposition of immunoglobulin fraction in the presence of 12% of polyethylene glycol. The precipitate immunoglobulins diluted in 0.9% sodium chloride solution, centrifuged to remove insoluble impurities. To the solution after centrifugation add Guetta, he provides a drug containing impurities of residual polyethylene glycol. In addition, the resulting solution of immunoglobulins contain significant amounts of fibrinogen and fibrin, shoesadidas with immunoglobulins. These impurities significantly complicate sterilizing filter and cause instability of the drug during storage in liquid form. The consequence of this is the need for immediate filling and freeze drying after filtration. Loss in the final release of the drug due to difficulties in filtration and the need to re-conduct this procedure reaches 30%.The problem solved by the invention is obtaining immunoglobulin preparation that is free of impurities volatile components, complicating filtering.The technical result from the use of the invention is to increase the yield of the target product and reducing its cost.This result is achieved in that in a method of producing immunoglobulin preparation from human blood by salt extraction of the alcohol precipitate serum, clearing the lipoproteins, followed by separation of zelenoklyniv conduct alcohol precipitation, as a stabilizer, a mixture of glycine and glucose at a ratio (1-1,5):1, a solution of immunoglobulins treated sorbents made of glass, at the stage of selection of the target product are conducting a flow-zonal centrifugation, and before sterilizing pass filtering the solution through a porous glass filter.The method is as follows. Deposition of immunoglobulin fraction of the chloroform extract of the precipitate B is carried out with the use of ethyl alcohol. This avoids the presence in the final product of polyethylene glycol, which according to the method prototype must be removed by repeated washing. Alcoholic residue is dissolved in 0,55 - 0,65% solution of sodium chloride with the addition of glycine and glucose at a ratio (1-1,5):1 (i.e. 20.5% of glucose and 2-3% glycine), pH 6.5-7.5. These values of pH and salt are the best because they provide the most complete formation of a precipitate of insoluble fibrin, and do not require further correction. Reducing the salt concentration below 0,55% impedes the dissolution of the sediment, and increase more of 0.65% makes it difficult subsequently held the lyophilization. The increase in the ratio of the concentrations of glycine and glucose does not increase the yield of the final PR is of fibrin sediment spend processing solution using glass beads. The solution is incubated for about 30 minutes until the formation of fibrin, then subjected to flow-zonal centrifugation at 15000 rpm. /min. and the Supernatant passed through a filter of porous glass (filter Shota) in order to remove residual amounts of fibrinogen and fibrin, adsorbed on porous glass. The resulting solution was subjected to sterilizing filtration. This solution is stable and can be stored for up to filling in for 1 month without changing the physical-chemical properties (transparency, lack of deposited components).Example 1. To 1 kg of sediment B add 10 l of 0.9% sodium chloride solution with stirring. The volume of extract 10 l protein 2,00,2%. To extract sediment B add 10% chloroform by volume. The mixture is slowly stirred for 2 hours the Precipitate is removed by centrifugation. Set the pH of the solution is equal to 6,90,5. Add ethanol to a final concentration of 26%. The precipitate was separated by centrifugation at 15000 rpm./minutes the precipitate immunoglobulins diluted in 0,60,05% solution of sodium chloride with the addition of 20.5% glucose and 3.0% glycine. To the solution add glass beads and incubated for about 30 minutes until the formation of fibrin. After processing, carried out the separation of the fibrin Ecay through the filter Shota. Then a solution of immunoglobulins is subjected to sterilizing filtration using a depth cellulose filters Cuno-60" and "Cuno-90", and lyophilized using the apparatus of TGS -15.Specific activity (titer) of the obtained product is not different from the prototype (Shigella - 1:320, Salmonella 1:320 to 1:640). The content of immunologically inactive fractions, determined by electrophoresis on membranes from acetylcellulose, also does not differ from the prototype and is 0-1%. The output of the immunoglobulin preparation is 170-185 doses of 1 kg of raw material.Example 2. Carried out analogously to example 1, but adding glycine to 2%. The final output of the product - 170-185 doses of 1 kg of raw materials, specific activity - titer antibodies to Shigella - 1:320, Salmonella - 1:1320 content immunologically inactive fractions, determined by electrophoresis on membranes from acetylcellulose, - 1%.Use as precipitating reagent instead of glycol ethyl alcohol can improve the output of the immunoglobulin fraction by 15%, increase speed centrifugation also gives an increase in the yield of immunoglobulins by 15%. The removal of impurities fibrin leads to reduce the d drug is 170-185 doses of 1 kg of raw materials, while the method of the prototype allows you to receive no more than 100 doses of 1 kg of raw material. The total increase in the yield of the product is 70-85%. The cost of the drug compared with the prototype reduced by 50%. The method of obtaining the immunoglobulin preparation from human blood by salt extraction of the alcohol precipitate serum, clearing the lipoproteins, followed by separation of the target product, adding a stabilizer, containing glucose, sterilizing filter, characterized in that the selection of immunoglobulins conduct alcohol precipitation, as stabilizer, a mixture of glycine and glucose in the ratio (1 to 1.5) : 1, treated with a solution of immunoglobulins sorbents made of glass, at the stage of selection of the target product are conducting a flow-zonal centrifugation, and before sterilizing filtration miss the immunoglobulin solution through a porous glass filter.
FIELD: genetic engineering, immunology, medicine.
SUBSTANCE: invention relates to new antibodies directed against antigenic complex CD3 and can be used in therapeutic aims. Antibody IgG elicits the affinity binding with respect to antigenic complex CD3 wherein heavy chain comprises skeleton of the human variable region in common with at least one CD3 taken among amino acid sequences SEQ ID NO 2, 4 and 6 and their corresponding conservatively modified variants. Light chain comprises skeleton of the rodent variable region in common with at least one CD3 taken among amino acid sequences SEQ ID NO 8, 10 and 12 and their corresponding conservatively modified variants. Antibody is prepared by culturing procaryotic or eucaryotic cell co-transformed with vector comprising recombinant nucleic acid that encodes antibody light chain and vector comprising recombinant nucleic acid that encodes antibody heavy chain. Antibody is administrated in the patient suffering with malignant tumor or needing in immunosuppression in the effective dose. Invention provides preparing chimeric antibodies against CD3 that are produced by expression systems of procaryotic and eucaryotic cells with the enhanced yield.
EFFECT: improved preparing methods, valuable medicinal properties of antibody.
33 cl, 5 dwg, 1 ex
FIELD: medicine, pharmaceutical industry and technology, pharmacy.
SUBSTANCE: invention relates to a composition eliciting an antiviral effect. The composition comprises hydrophilic conglomerate of immunoglobulins consortium adsorbed with polyethylene glycol 4000-6000, recombinant interferon-α2 and a special additive taken among the following substances: glycine, glucose, maltose, sodium chloride taken in the definite ratio of components. Invention provides elevating solubility of composition eliciting an antiviral effect and enhanced release of biologically active substances to solution.
EFFECT: valuable medicinal properties of composition.
FIELD: medicine, pharmacy.
SUBSTANCE: invention relates to a composition eliciting an antibacterial effect. Composition comprises hydrophilic conglomerate of immunoglobulins consortium adsorbed with polyethylene glycol 4000-6000 and a special additive taken among the following substances: glycine, glucose, maltose, sodium chloride taken in the definite ratio of components. Invention provides sufficient desorption of biologically active substances in resuspending the composition eliciting an antibacterial effect and comprising consortium of immunoglobulins.
EFFECT: valuable medicinal properties of composition.
SUBSTANCE: the innovation deals with new immunogenic conjugates of beta-propionamide-bound polysaccharide and N-propionamide-bound oligosaccharide with protein, and the method to obtain these conjugates has been suggested, as well. Conjugates should be applied to obtain vaccines against infectious diseases and cancer that enables to broaden the number of preparations applied in treating the above-mentioned diseases.
EFFECT: higher efficiency.
1 dwg, 2 ex, 8 tbl
FIELD: microbiology and immunology, in particular immunodiagnosis.
SUBSTANCE: atypical strain of melioidose Burkholderia pseudomallei-111-6-1 with altered phenotype defected with respect to synthesis of 8 antigen and acting as immunosuppressor is used as antigen for animal immunization. Immune serum is obtained after 2 immunization cycles of animal-producer with titer in gel immunodiffusion reaction not less than 1:128.
EFFECT: immune serum with increased specific activity.
2 tbl, 2 ex
FIELD: medicine and immunology, in particular treatment and prevention immunodeficiency conditions and diseases associated with bacterial or viral aggression.
SUBSTANCE: claimed method includes administration to a patient immunoglobulin drug (e.g., pharmaceutical composition containing 6-12 % of specific heterologous secreted immunoglobulin A, isolated from milk or foremilk of immunized ungulates). Administration is performed parenterally wherein single dose is at least 10 IU/kg of patient weight for treatment or at least 5 IU/kg for prophylaxis; or perorally in dose of 0.2-0.5 g and/or topically one-two times per day for 1-5 days. Method of present invention makes it possible to decrease dose of administrating immunoglobulin due to prolonged retention of its high titers in body fluids.
EFFECT: enlarged range of application and assortment of immunoglobulin drugs.
4 cl, 5 ex
SUBSTANCE: the present innovation deals with cryoprotective ointment containing recombinant interferon-α2. The suggested cryoprotective ointment contains recombinant interferon-α2, glycerol, polyethylene glycol 300-6000, polyglucin, buffered 0.02%-Trilon B solution at pH of 5.5-7.0 and ointment foundation at a certain content of components per 1.0 g ointment. Additionally, cryoprotective ointment could contain glycine 3,7-bis(dimethylamino)phenothiazonium chloride, dry immunoglobulin preparation or dry immunoglobulin preparation for enteral application. Ointment foundation of cryoprotective ointment could contain water-free lanolin, Vaseline and Vaseline oil, at the following ratio of components: 2.5;3.5:1 - 6.5:0.5:1. The innovation provides maximal safety of recombinant interferon-α2 activity in cryoprotective ointment at multiple alteration of positive and negative environmental temperature and at keeping cryoprotective ointment under these conditions.
EFFECT: higher efficiency of application.
8 cl, 8 ex
FIELD: medicine, pharmaceutics, pharmacology.
SUBSTANCE: one should apply mammalian anti-HBP-antibodies. The ways are being suggested to identify monoclonal antibody bound, at least, with one epitope upon native HBP (heparin-binding protein) and methods to detect whether a mammal produces HBR being bound with a monoclonal antibody and, also, the kits for the above-mentioned purpose. The present innovation provides the opportunity to apply the mentioned antibodies in preventing and treating disorders associated with bradykinin releasing.
EFFECT: higher efficiency of application.
25 cl, 11 dwg, 3 ex, 1 tbl
FIELD: veterinary science.
SUBSTANCE: animals should be introduced with antihistamine serum (AHS) subcutaneously at the dosage of 4.0-5.0 ml in combination with myxoferon at the quantity of 60-75 dosages and vitamin C at the dosage of 1.0-1.5 ml/animal, once daily, thrice at interval of 5-7 d. Application of AHS in combination with myxoferon and ascorbic acid provides active stimulation of immunological reactivity, increases total body resistance I animals and causes no toxic effects and allergic reactions.
EFFECT: higher efficiency of correction.
FIELD: immunology, biotechnology, medicine.
SUBSTANCE: invention relates to antiidiotypical monoclonal antibody or fragment thereof for BSW17 antibody effecting on LgE Cε3-region bonding to high affinity LgE receptor. Amino acid sequence is as described in specification. antiidiotypical antibody is useful as pharmaceutical composition ingredient for LgE-mediated disease treatment. Invention make in possible to prevent allergic disorders and inflammations due to inhibiting interaction between LgE Cε3-region with high affinity receptor by claimed antibody.
EFFECT: new agent for allergic and inflammation disorder treatment.
7 cl, 32 dwg, 5 tbl, 10 ex