The method of producing drug for indication of the toxin of diphtheria pathogens in a liquid nutrient medium
(57) Abstract:A new method of producing drug for indication of diphtheria toxin in liquid nutrient medium. The method is carried out in 0.05 M glycine-saline buffer solution with a pH of 8.2 - 8,25 by immobilization of gammaglobulin, immune to the toxin of diphtheria pathogens on the surface of particles of polystyrene latexes activated methacrylic acid and methacrylate zinc, in the presence of bovine serum albumin, polyethylene glycol and preservatives. Used latex is a mixture of modified polystyrene latexes with particle size of 0.20 - 0.25 μm and 1.1 to 1.2 μm. Components for the production of drug taking in a certain weight ratio. The inventive method relates to the field of Microbiology, and in particular to a drug necessary for carrying out immunological analysis to indicate the toxin of diphtheria pathogens.Specified drug is needed to identify toxinogenesis the causative agent of diphtheria patients and carriers, to monitor the effectiveness of treatment of patients with diphtheria and sanitation carriers of the causative agent of diphtheria, for assessing the efficacy of preventive and protivoradarnyh articles Mazurova I. K., Kombarov C. Yu, L. Spring M, Grigorian, K. Reaction of coagglutination for the detection of diphtheria toxin. Journal of Microbiology, epidemiology and Immunobiology". - 1989 - N 3 C. 68-74; Osipova, F., Barashkova L. N. Determination of toxigenic properties corynebacteria diphtheria in the reaction of indirect hemagglutination with the use or antibody-based test diagnostic drug. Abstr. Novgorod, 1989. Topical issues of infectious diseases and its prevention in the Novgorod region. 8th conference of the epidemiology, Microbiology, Parasitology. - S. 54-56; Mazurov, I. K., Potemkin, E. E. , Sviridov centuries, Zaitsev, E. M. Detection of toxin and the assessment of the degree of coccinobaphi Corynebacterium diphitheriae using enzyme immunoassay. "The journal of Microbiology, epidemiology and Immunobiology", - 1989 N 6 C. 66-70).In the first drug as a carrier of antibodies used protein And Staphylococcus, which complicates the technology of preparation of the drug and does not provide receive high quality product. The second drug used in the reaction of indirect haemagglutination, is a sheep erythrocytes loaded with antibodies to diphtheria toxin. This drug provides good sensitivity of the method, but obtaining the purified antibodies to diphtheria toxin, adsorbed on the surface of special tablets. Immunoassay method has high sensitivity, but costly, complex staging, requires special equipment and trained personnel.The prototype of the proposed method is latex preparation, previously proposed by the authors for the identification of streptococcal groups A, B, C, D and described in applications NN 92008090, 92008091, 92008092 of 26 November 1993, the Essence of the prototype is immobilization gamma globulin, immune to Streptococcus, on the surface of latex particles, which are particles of polystyrene of different dimensions and activated methacrylate zinc.The technical essence of the proposed method is that the target of the drug is produced by sorption of highly purified, immune to diphtheria toxin antibodies on the latex particles produced from polystyrene modified with methacrylic acid and methacrylate zinc. The process includes the following operations.1. The mixture of antibody-gammaglobulin, immune to diphtheria toxin diluted in 0.05 M glycine-saline buffer solution with a pH of 8.2, latex, diluted with the same buffer solution to the latex content in the buffer solution of 1%m zinc with surround enabled dispersion 1,1 - 1.2 µm and latex polystyrene carboxylating with a dispersion of 0.2 - 0.25 μm. The volumetric ratio of these latexes in the mixture is 1:2.2. Received on p. 1 mixture incubated at 56oC for 45 minutes3. The mixture obtained under item 2, cool to 4 - 6oC.4. The cooled mixture is mixed with bovine serum albumin, pre-diluted with 0.05 M glycine-saline buffer solution with a pH of 8.2 to a concentration of 0.1%.5. To the mixture obtained under item 4, enter the polyethylene glycol-6000 and preservative.6. The components necessary to prepare the target of the drug, taken in the following ratio in g/l:
gammaglobulin, immune to diphtheria toxin - 0,05 - 0,06
the latex mixture - 2,50 - 2,55
bovine serum albumin - 0,5 - 0,55
the polyethylene glycol-6000 - 55,0 - 60,0
preservative - 0,1 - 1,0
0.05 M glycinato-salt buffer solution with a pH of 8.2 - 8,25 - Rest
The inventive method is simple and provides highly sensitive drug.The authors argue that the claimed method meets the criteria of the invention "inventive step", since on the basis of acquaintance with scientific, technical and patent information, and twitcam from the prior art, related to the declared object.The inventive method is carried out as follows.1. Gammaglobulin, immune to diphtheria toxin, plant 0.05 M glycine-saline buffer solution with a pH of 8.2 to a concentration of 0.25 mg/kg2. Polystyrene latex, activated methacrylate zinc, with a particle size of 1.2 μm bred 0.05 M glycine-saline buffer solution with a pH of 8.2 to a concentration of 1% (based on the polymer.3. Polystyrene carboxypropanoyl latex with a particle size of 0.2 μm bred 0.05 M glycine-saline buffer solution with a pH of 8.2 to a concentration of 1% (based on the polymer.4. Prepared by PP. 2 and 3 polystyrene latexes take in a volume ratio of 1:2.5. 10 ml of the composition obtained under item 1, incubated with 10 ml of latex suspensions, obtained under item 4, in a water bath at 56oC for 45 minutes6. The mixture obtained under item 5, cooled to 4 to 6oC for 18 - 20 h, add 20 ml of 0.1% solution of bovine serum albumin in 0.05 m glycine-saline buffer solution with a pH of 8.2, containing 12% peg-6000. In the resulting mixture was added 0.004 g of thimerosal (preservative).The results of operations item is manufactured in the above way, it has a high specificity, which is manifested in the absence of cross-reactions against other antigens toxins secreted by pathogens other diseases.The results of sensitivity studies of the drug received in response latex agglutination (RLA) presents test RLA on glass with diphtheria toxoid - positive at a concentration of 10-3- 10-4Lf/mlNote: Lf - unit flocculation toxin causative agent of diphtheria. The method of producing drug for indication of the toxin of diphtheria pathogens in a liquid nutrient medium under conditions of glycine-saline buffer solution with a pH of 8.2 to 8.25 by immobilization of gammaglobulin, immune to diphtheria toxin, particles of polystyrene latex, including stabilization in the presence of bovine serum albumin, polyethylene glycol-6000 and preservative, characterized in that the process is carried out in glycine-saline buffer solution with both molarity of 0.05, as polystyrene latex, a mixture of polystyrene latex with a particle size of 1.1 to 1.2 μm, activated methacrylate zinc when surround is enabled, and carboxylating polystyrene latex with a particle size of 0.2 to 0.25 m The latex mixture 2,5 2,55
Bovine serum albumin 0,5 0,55
The polyethylene glycol-6000 55 60
Preservative 0,1 1,0
0.05 M glycine-saline buffer solution with a pH of 8.2 to 8.25 Ostalnoe
FIELD: medicine, oncology, immunology, tumor biology.
SUBSTANCE: invention relates, in particular, to methods for enhancing cytotoxicity based on applying anti-CD38-immune toxins. Method involves carrying out the treatment of patient with pathophysiological state taken among the group including myelomas and leukosis and involves the following stages: a) administration to the indicated patient the pharmacologically effective dose of retinoid that enhances expression of antigen CD38; and b) administration to the indicated patient the pharmacologically effective dose of immune toxin acting against effectively expressing antigen CD38. Method provides enhancing the cytotoxicity with respect to above said diseases in their resistance to anti-tumor medicinal agents.
EFFECT: enhanced and valuable method for treatment.
6 cl, 1 tbl, 9 dwg, 10 ex
SUBSTANCE: method involves applying effective doses of epidermis growth factor receptors antagonists being active ingredient in drug for inhibiting various types of resistant human tumors. The epidermis growth factor receptor antagonists are combined with other chemotherapeutic agents like Cisplatin, Irinotecan or ionizing radiation.
EFFECT: enhanced effectiveness of treatment.
50 cl, 2 tbl
FIELD: medicine, molecular biology, antibodies.
SUBSTANCE: invention relates to an antibody raised against CCR5 and comprising: (i) two light chains wherein each light chain comprises product of plasmid expression and designated as pVK:HuPRO140-VK (ATCC - PTA-4097), and (ii) two heavy chains wherein each heavy chain comprises product of plasmid expression and designated as pVg4:HuPRO140 HG2-VH (ATCC - PTA-4098), or plasmid designated as pVg4:HuPRO140 (mut B+D+I)-VH (ATCC - PTA-4099), or fragment of such antibody binding with CCR5 on a human cell surface. Invention relates to nucleic acid encoding light and heavy chains of antibody, expression vector, cell-host transformed with at least one vector, and a method for preparing antibody. Antibody is used as an active component in composition used for inhibition of infection of cells CD4 + HIV-1, and to a pharmaceutical composition used in treatment of a patient with HIV-1 infection. Also, invention relates to antibody conjugate against CCR5 and its using. Use of antibodies provides enhancing effectiveness of prophylaxis and treatment of HIV-1 infection.
EFFECT: valuable medicinal properties of antibody.
31 cl, 23 dwg, 3 ex
FIELD: medicine, oncology.
SUBSTANCE: method includes the consecutive stages: (a) administration of at least one dose of anti-angiogenic cyclo-(arginine-glycine-asparagine acid)-containing pentapeptide (pentapeptide cRGD), such as cyclo-(Arg-Gly-Asp-D-Phe-[N-Me]-Val); (b) administration of anti-tumor effective amount of radio immunotherapeutic agent(RIT) not later than in 1 hour following administration of pentapeptide cRGD at stage (a); and (c) administration of at least two additional doses of pentapeptide cRGD, where the first additional dose is administered within 2 days after RIT and each additional dose of pentapeptide cRGD is administered with intervals between doses not more than 2 days.
EFFECT: invention provides the synergic effect in regard to apoptosis of tumor cells and endothelial cells of tumor vessels.
30 cl, 6 dwg, 2 tbl
FIELD: medicine; immunology.
SUBSTANCE: sorbent is offered to remove immunoglobulin from human blood plasma. This sorbent contains agarous matrix covalently combined with ligand. As a ligand at that it contains F(ab)2 fragments of specific affinely-purified polyclonal antibodies blocking human immunoglobulin G. Sorbent is actually biologically inert, biocompatible agarous matrix. Sorbent is characterized with higher sorptive capacity and safety of immunosorbents used practical purposes, specifically for therapeutic aphaeresis in comparison to well-known polyclonal bodies based sorbents.
EFFECT: considerable reduction of prospective immunological response of human body for foreign protein.
1 ex, 1 tbl
FIELD: medicine; oncology.
SUBSTANCE: invention can be used for treatment of an acute myelogenetic leukemia or myelodysplastic syndrome. For this purpose use a combination of preparations hemetuzumab ozohamicin, daunorubicin and cytarabinum in certain doses and regimens.
EFFECT: invention promotes effective treatment of the specified diseases due to synergistic effect at influence of these preparations on an organism.
2 cl, 2 ex
SUBSTANCE: invention refers to medicine, namely to ophthalmology, and can be used in treating the patients with macular oedemas of various geneses. That is ensured by the therapy that starts with 6 sessions of plasma exchange every 2 days. It is followed by a single intravitreal introduction of antivasoproliferative preparation Avastin in dosage 1.25 mg or Lucentis in dosage 0.5 mg.
EFFECT: method allows improving clinical effectiveness combined with reduced intravitreal introduction of the antivasoproliferative preparations, due to involving pathogenetic mechanisms, namely elimination of pathological immune complexes, stabilisation of pro- and anti-inflammatory cytokines.
2 cl, 2 ex
SUBSTANCE: copolymers of hetero-chain aliphatic poly-N-oxides of general formula (I) , where R=N, CH; x=2-4; y=0, 2; n=10-1000; q=(0.1-0.9)n; z=(0.1-0.9)n. Copolymers possess anti-oxidant action, therapeutic action as detoxicant and immunomodelling agent. Copolymers of formula (I) can be used as immunomodulating carrier for obtaining vaccinating medication and as carrier of medications for obtaining medications.
EFFECT: copolymers of hetero-chain aliphatic poly-N-oxides represent novel class of compounds possessing wide spectrum of pharmacological and vaccinating action, aimed at increase of safety in application, increase of technological and economical effectiveness and ecological safety of production of medications.
20 cl, 2 dwg, 13 tbl, 22 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology and represents methods of treating an autoimmune disease involving administering into an individual a pharmaceutical composition containing a pharmaceutically acceptable carrier and a humanised anti-CD4-antibody, able to activate the regulatory C-cells CD4+CD25+, wherein the above antibody contains V-domain of H-chain containing the sequences DCRMY, VISVKSENYGANYAESVRG and SYYRYDVGAWFAY, and V-domain of L-chain containing the sequences RASKSVSTSGYSYIY, LASILES and QHSRELPWT; the above composition is administered subcutaneously into the individual in a dose of the humanized anti-CD4-antibody 20 to 200 mg or 8 to 60 mg/m2 of the individual's body surface, or 0.2 to 2 mg/kg. The present invention also discloses the pharmaceutical compositions applicable in the above methods of treating the autoimmune disease.
EFFECT: invention enables administering the pharmaceutical composition containing the humanised anti-CD4-antibody in higher doses losing no therapeutic effect and causing no intensification of any side effects.
46 cl, 20 dwg, 21 tbl, 8 ex
SUBSTANCE: invention represents methods for treatment of an autoimmune disease, including introduction of a pharmaceutical composition to a subject, containing a pharmaceutically acceptable carrier and a humanised anti-CD4-antibody, capable of activating regulatory T-cells CD4+CD25+, where the specified antibody contains a V-domain of an H-chain, containing sequences DCRMY, VISVKSENYGANYAESVRG and SYYRYDVGAWFAY, and a V-domain of an L-chain, containing sequences RASKSVSTSGYSYIY, LASILES and QHSRELPWT. The specified composition is introduced to a subject with frequency from daily intake to introduction every 31st day and with a dose of a humanised anti-CD4-antibody from 20 to 200 mg, or from 8 to 60 mg/m2 of the subject body surface area, or from 0.2 to 2 mg/kg. This invention also discloses a set for treatment of an autoimmune disease, which contains multiple doses of the above pharmaceutical composition.
EFFECT: invention makes it possible to introduce a pharmaceutical composition containing a humanised anti-CD4-antibody, with more lengthy intervals of dosing and in higher doses, not losing the therapeutical effect and not causing side effects.
59 cl, 41 dwg, 27 tbl, 9 ex