The method of obtaining igm-containing concentrate of immunoglobulin
(57) Abstract:The invention relates to medicine and for the production of therapeutic drugs from plasma or serum of donor blood, namely, the method of obtaining lgM-containing concentrate of immunoglobulin. The method consists in the fact that the precipitate II+III, obtained by ethanol fractionation of plasma (serum) blood is processed sequentially by the Aerosil-380 and sodium capricolum with intermediate zentrifugenbau removal of ballast proteins, subjected to ultrafiltration to remove low molecular weight impurities and concentration. After sterilizing filtration obtain the target product containing an average of 15% of total IgM immunoglobulin. The advantage of the invention consists in the enrichment of immunoglobulin IgM. table 1. The invention relates to medicine and relates to a therapeutic drug from plasma donated blood
Raw material for the production of the drug is plasma donors tested for the absence of the surface antigen of hepatitis B virus (HBsAg), antibodies to human immunodeficiency virus (HIV) and hepatitis C virusThe prototype of the invention is a method of producing immunoglobulin is aceski immunoglobulin contains mainly immunoglobulin G class, which greatly limits its activity against gram-negative microorganisms.The aim of the present invention to provide IgM-containing concentrate of immunoglobulin.The method is as follows. Combined pool of plasma fractionary ethanol in the cold to the stage of precipitate II+III; crude residue is suspended in 0.9% sodium chloride in a weight ratio of 1:10 and treated with Aerosil-380, ballast sediment removed by centrifugation at 13,000 g; add sodium kapalbekuly to a final concentration of 0.4% at slightly acidic pH values; conduct maturation of the drug for the formation of a precipitate unstable proteins, which are removed by centrifugation; perform ultrafiltration to remove low molecular weight impurities and concentration and after sterilizing filtration obtain the target product.Example. Immune plasma in the amount of 360 l obtained by the method of plasmapheresis. Spent the alcohol fractionation at low temperatures. Received 21000 g wet sediment fraction II+III. For the preparation of IgM-containing concentrate of immunoglobulin 260 g of sediment suspended under stirring in 2600 ml of 0.9% restr>C for 18 hours Aerosil-380. After 1 hour, Aerosil and ballast proteins were removed by centrifugation at 13,000 g. The volume of centrifugate was 2170 ml of 0.06 M acetate buffer brought the pH of centrifugate to 4.0, added kaprilat sodium to a final concentration of 0.4%. After maturation at a temperature of (203)oC for 18 hours spent centrifugation at 13,000 g. Then spent diafiltration of centrifugate 5-fold volume of sodium chloride 0.9 per cent, the ultrafiltration to a protein concentration of 5.3%, was added to the concentration of glucose (1,00,2)%, glycol to a concentration of (0,50,2)%. Spent sterilizing filtration of the solution. Received 200 ml of concentrate. The concentrate of immunoglobulin poured in bottles of 50 ml and 25 ml of the target product.In laboratory preparations of blood of the Kirov research Institute of Hematology and blood transfusion of the developed technology made 7 series concentrate. The table below summarizes the results of the evaluation of physicochemical and biologic properties of the obtained series. The concentrate produced a clear, colorless or slightly opalescent solution, transparency averaged (0,0130,001), chromaticity (0,046 0,002) units). the density. During the observation period (up to 12 months.) education is Vila average (5,67 of 0.07)%.Compared with the prototype concentrate has a low anticomplementary activity (AKA): in the experimental samples it ranged from 3.2 to 6,54 mg protein, not activating 2 CH50the complement. (Level AKA intramuscular immunoglobulin differs from that of figure 3 of the order and approximately 1,510-3mg).Evaluation of the fractional composition of the samples obtained by the method of immunoelectrophoresis revealed the presence of 4 arcs of precipitation corresponding to IgG, IgA, IgM, and a small quantity of albumin. The percentage of IgG:IgA:IgM was equal to the average of 66.6:18,1:15,3%.Molecular parameters of the samples was estimated by the method of gel chromatography of low pressure with an ACA-34. Elution was performed 0.05 M phosphate buffer. The chromatogram was allocated 3 peak corresponding to the fractions with molecular masses of more than 160 thousand D (IgM), 150-160 thousand D (total peak IgG and IgA), the third - 50-70 thousand Days Last indicate the presence of albumin. The absence in samples of polymers IgG was confirmed by the absence in the faction more than 160 thousand D immunoglobulin G, is proved by the method of radial immunodiffusion according to Mancini. The percentage fractions was as follows: the content of IgM ranged from 9 to onsite, aerogenes.According to the TPHA concentrate has a high activity of antibodies to Pseudomonas aeruginosa: the average geometric return titer was 146,9. According to ELISA the presence of IgG antibodies to lipopolysaccharides of Pseudomonas aeruginosa ranged from 3,28 to 16,58 EDELISA/mg protein, IgM-antibodies - from 0.18 to 1.80 EDELISA/mg protein. The corresponding figures for E. coli averaged 8,31 of 0.85 and 1.12 of 0.25. EDELISA/mg protein. It must be emphasized that in the preparation of immunoglobulin produced by the method-prototype, reverse titer antiintegrin antibodies in TPHA hovers around 80, ELISA activity of IgG to LPS Escherichia coli is equal to the average of 1.60 0,27, IgM - 0,01 0,01; to E. coli - 10,46 0,87, IgM - 0,03 0,01 UELISA/mg protein.Thus, the obtained concentrate immunologically active fraction of donor blood containing an average of 15% of total IgM immunoglobulin and highly active antibodies against gram-negative pathogens and low anticomplementary activity.The proposed method of producing IgM-containing concentrate of immunoglobulin from the precipitate fraction II+III can be used when creating the production technology IgM-containing drug immunodeficiency is the conduct alcohol plasma fractionation at low temperatures, get the crude residue fraction II + III, characterized in that it is suspended in 0.9% sodium chloride in a weight ratio of 1 : 10 and treated with Aerosil-380, ballast sediment removed by centrifugation at 13000g, add sodium kapalbekuly to a final concentration of 0.4% at slightly acidic pH values, conduct maturation concentrate for the formation of a precipitate unstable proteins, which are removed by centrifugation, perform ultrafiltration to remove low molecular weight impurities and the concentration of the drug and after sterilizing filtration obtain the target product.
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