Antibodies and kin molecules bound to psca proteins
SUBSTANCE: invention is related to biotechnology and represents monoclonal antibody or its antigen-binding fragment, containing antigen-binding site, which specifically binds with protein of prostatic stem cells antigen (PSCA). At the same time monoclonal antibody is produced with postfusional cell line, selected from group of postfusional cell lines deposited in American typical cultures collection (A.T.C.C.) under inventory No.PTA-6698, PTA-6699, PTA-6700, PTA-6701, PTA-6702 and PTA-6703. Besides invention is also related to expression vector, which contains polynucleotide coding this antibody, and also to method for analysis for detection of PSCA protein presence in biological sample with application of this antibody. Moreover, invention is related to method for delivery of cytotoxic agent or diagnostic agent to cell, which expresses PSCA protein, with application of this antibody, and also to method for detection of PSCA protein in biological sample with application of above-mentioned antibody.
EFFECT: invention may efficiently be used in active and passive immunisation against cancer diseases.
21 cl, 27 dwg, 11 tbl, 33 ex
The text descriptions are given in facsimile form.
1. Monoclonal antibody or its antigennegative fragment containing antigennegative website, which is specifically associated with protein antigen prostatic stem cells (PSCA) (SEQ ID NO: 2), where a monoclonal antibody is produced by hybridoma cell line selected from the group hybridoma cell lines deposited at the American type culture collection (ATIS) under inventory No. of MOUTH-6698, MOUTH-6703, MOUTH-6699, MOUTH-6700, MOUTH-6701 and MOUTH-6702.
2. Monoclonal antibody or its antigennegative fragment according to claim 1, where the monoclonal antibody is produced by hybridoma cell line under inventory No. ASS MOUTH-6701.
3. The antibody or antigennegative fragment comprising antigennegative site that binds with the protein (PSCA) (SEQ ID NO: 2), where antigennegative site includes the amino acid sequence of variable region of the heavy chain and the variable region of the light chain of the antibody secreted hybridoma cell line selected from the group of cell lines deposited in ASS under inventory No. of MOUTH-6698, MOUTH-6703, MOUTH-6699, MOUTH-6700, MOUTH-6701 and MOUTH-6702.
4. Monoclonal antibody or its antigennegative fragment according to claim 3, where the hybridoma cell line is a line under inventory No. ASS MOUTH-6701.<> 5. The antibody or antigennegative fragment containing antigennegative site which specifically binds to PSCA protein (SEQ ID NO:2), where the monoclonal antibody includes the amino acid sequence of the variable region of the heavy chain (VH) of SEQ ID NO: 22 and the variable region of the light chain (VL) of SEQ ID NO: 23; a VH sequence SEQ ID NO: 17 and a VL sequence SEQ ID NO: 18; a VH sequence SEQ ID NO: 13 and a VL sequence SEQ ID NO: 14; a VH sequence SEQ ID NO: 15 and a VL sequence SEQ ID NO: 16; a VH sequence SEQ ID NO: 19 and a VL sequence SEQ ID NO: 21; or a VH sequence SEQ ID NO: 28 and a VL sequence SEQ ID NO: 29.
6. The antibody or fragment according to any one of claims 1 to 5, where the fragment is a Fab fragment, F(ab')2, Fv or Sfv.
7. The antibody or fragment according to any one of claims 1 to 6, where the antibody or fragment is conjugated with detektivami marker, a toxin, a therapeutic agent or chemotherapeutic agent.
8. The antibody or fragment according to claim 7, where the detected marker is a radioisotope, a metal chelator, an enzyme, fluorescent compound, bioluminescent compound or a chemiluminescent compound.
9. The antibody or fragment of claim 8, where the radioisotope include212Bi131I131In90Y186Re,211At,125I188Re,153Sm213Bi32P is Li Lu.
10. The antibody or fragment according to claim 7, where the toxin include ricin, domain a of ricin, doxorubicin, daunorubicin, maytansinoid, Taxol, ethidium bromide, mitomycin, etoposide, teniposide, vincristine, vinblastine, colchicine, dihydroxyanthracene, actinomycin, diphtheria toxin, exotoxin A of Pseudomonas (RE), RE, abrin, domain And abrina, domain And modeccin, alpha sarcin, gelonin, mitogillin, restrictocin, vanomycin, inomycin, kouritzin, krotin, calicheamicin, inhibitor Saponaria officinalis, glucocorticoid, auristatin, aureomycin, yttrium, bismuth, complestatin, duocarmycin, dolastatin, SS or cisplatin.
11. Hybridoma producing a monoclonal antibody according to claim 1 or 2, where hybridoma selected from the group consisting of lines under inventory No. of MOUTH-6698, MOUTH-6703, MOUTH-6699, MOUTH-6700, MOUTH-6701 and MOUTH-6702.
12. The expression vector containing polynucleotide encoding the antibody comprising the amino acid sequence of the variable region of the heavy chain (VH) of SEQ ID NO: 22 and the variable region of the light chain (VL) of SEQ ID NO: 23; a VH sequence SEQ ID NO: 17 and a VL sequence SEQ ID NO: 18; a VH sequence SEQ ID NO: 13 and a VL sequence SEQ ID NO: 14; a VH sequence SEQ ID NO: 15 and a VL sequence SEQ ID NO: 16; a VH sequence SEQ ID NO: 19 and a VL sequence SEQ ID NO: 21; or a VH sequence SEQ ID NO: 28 and a VL sequence SEQ ID NO: 29.
13. The vector according to item 12, which is olignucleotides encodes a single-chain antibody.
14. The method of analysis for the detection of the presence of PSCA protein in a biological sample, comprising bringing the sample into contact with an antibody according to any one of claims 1 to 6, and the detection of binding protein PSCA (SEQ ID NO: 2) in the sample.
15. The method of delivery of cytotoxic tools or diagnostic agent to a cell expressing PSCA protein (SEQ ID NO: 2), including:
ensuring cytotoxic tools or diagnostic agent conjugated to an antibody or its fragment according to any one of items 1 to 6, with the formation of the conjugate of the antibody-agent or fragment-antibody funds; and
the effect on the cell with a conjugate of the antibody-agent or fragment of antibody-tools.
16. The method according to clause 15, where the cytotoxic agent or a diagnostic agent selected from the group consisting of a detected marker, toxin and remedies.
17. The method according to clause 16, where the detected marker is a radioisotope, a metal chelator, an enzyme, fluorescent compound, bioluminescent compound or a chemiluminescent compound.
18. The method according to 17, where the radioisotope include212Bi131I131In90Y186Re,211At,125I188Re,153Sm213Bi32P or Lu.
19. The method according to clause 16, where the toxin include ricin, domain a of ricin, doxorubicin, daunorubicin, maytansinoid, Taxol, ethidium bromide, mitomycin,etoposide, teniposide, vincristine, vinblastine, colchicine, dihydroxyanthracene, actinomycin, diphtheria toxin, exotoxin A of Pseudomonas (RE), RE, abrin, domain And abrina, domain And modeccin, alpha sarcin, gelonin, mitogillin, restrictocin, vanomycin, inomycin, kouritzin, krotin, calicheamicin, inhibitor Saponaria officinalis, glucocorticoid, auristatin, aureomycin, yttrium, bismuth, complestatin, duocarmycin, dolastatin, SS or cisplatin.
20. Method detection PSCA protein (SEQ ID NO: 2) in a biological sample, comprising the steps:
providing a biological sample and the control sample;
bringing into contact the biological sample and the control sample with the antibody according to any one of claims 1 to 6, which specifically binds to PSCA protein; and
determine the number of complex substances with the PSCA protein and antibody present in the biological sample and the control sample.
21. The method according to claim 20, further including:
the selection of the biological sample and the control sample from a patient that has cancer or suspected cancer of the prostate, pancreas, bladder, kidney, colon, lung, ovarian or breast cancer.
SUBSTANCE: invention can be used for production of monoclonal antibodies (MCAs) to heat shock protein 70 (HSP 70). A hybridoma strain is made by immunisation of BALB/c mice with bovine HSP 70 within 78 days. For the third days, splenocytes of immune mice (108 cells) are hybridised with murine myeloma cells P3-X63 Ag/8-653 (107 cells). A fusion agent is polyethylene glycol of molecular weight 4000 (Merk, Germany). The hybridisation is followed with selection, screening, cloning and cryopreservation of hybridoma. Hybridoma 6G2 is deposited in the microorganism collections of "ГНТТ ПМБ" under No. H-2. MCA.
EFFECT: produced hybridoma under the invention is more evident to be detected as HSP 70 on the cell surfaces, and change of endocellular HSP 70 level when exposed to the stress factors.
4 dwg, 1 tbl, 6 ex
SUBSTANCE: present invention refers to immunology and biotechnology. There is offered recovered human antibody to RG1 polypeptide. There are described versions of antibodies, including one-chain antibody, and immunoconjugate based on said antibodies. There are disclosed methods of selective cell destruction, cell inhibition, treatment of disease state, detection of disease state, detection of RG1, monitoring of clinical course of prostate cancer, prediction in a person with using antibodies and immunoconjugate.
EFFECT: application of the invention provides new antibodies to RG1 polypeptide that can find application in treating tumours with RG1 overexpression.
16 cl, 4 dwg, 1 tbl, 13 ex
SUBSTANCE: invention refers to antibody specifically getting bound with PRO87299 version. In addition, the antibody according to the invention has ability to block interaction HVEM and PRO87299 and to function as PRO87299 agonist. The antibody of agonist nature is produced by hybridoma Btig5F5.1 or Btig3B1.9. For the antibody, there is established amino acid sequence given in the description. The invention discloses the methods of using the antibodies to stimulate or reduction of immune response in immune-associated diseases connected, to relieve lymphoma, and inflammatory disease in requiring mammal, to detect polypeptide PRO87299 in a sample and to manage rejection of grafted cells.
EFFECT: antibody is an immunomodulator that allows applying therapeutically identical medicinal agents both to intensify and reduce immune response.
16 cl, 34 dwg, 7 tbl, 20 ex
SUBSTANCE: invention relates to Bcl-2 proteins, fragments thereof, and to application thereof in patients with a malignant tumour. The declared proteins and peptide fragments particularly are applicable in vaccine compositions for treatment of malignant tumour. Besides, the invention concerns the methods of treatment with application of specified compositions. Also, an aspect of the invention is production of T-cells and receptors thereof which are specifically recognise declared proteins and peptide fragments.
EFFECT: higher clinical effectiveness with respect to tumours.
61 cl, 5 ex, 2 tbl, 12 dwg
SUBSTANCE: invention concerns immunology area. Versions of the artificial fused protein consisting of an antibody (or its fragment) and cytokine, fused through a link peptide are offered. The antibody or its fragment is chosen from an antibody 225, 425, KS 1/4, 14.18, anti-CDx-antibody where x has the whole value 1-25. Each of versions of the fused protein has lowered quantity T-epitopes, at least, in the component of the fused protein presented by an antibody, and as consequence, possesses the lowered adjuvanticity, in comparison with an initial molecule. Identification of T-lymphocyte epitopes is performed by the automated calculation of sizes for the binding centres of class II MHC molecules with the subsequent experimental test of the obtained versions of protein for presence of the lowered adjuvanticity. The automated way of T-epitopes calculation is based on use of the Bjom's function modified in such manner that contribution of Van-der-vaals repulsion and lipophilic interaction in pairs between all lipophilic atoms of the chosen segments of the fused protein and a binding groove of a MHC P molecule is taken into account. Also a way of protein construction on the basis of the modified function Bjom's function with the subsequent experimental test of the received versions for presence of the lowered adjuvanticity is revealed, and also application of the fused protein for preparation of a pharmaceutical composition for tumour treatment is in addition considered.
EFFECT: invention use allows obtaining the fused proteins with the lowered adjuvanticity and, basically, keeping identical biological activity in comparison with a parent molecule; it can be used in treatment of tumours.
4 cl, 6 dwg, 22 tbl, 19 ex
SUBSTANCE: proposed is a recombinant single-strand trispecific antibody for treating tumours which express CEA. The said antibody consists of a series of three antibody fragments: anti-CEA-scFv, anti-CD3-scFv and VH CD28-antibody, linked by two intermediate linkers (intermediate linker Fc and intermediate linker HSA). If necessary, a c-myc-mark or (His)6-mark can be added at the C-end. Described is DNA, which codes the antibody, expression vector based on it and E.coli cell, containing the vector.
EFFECT: use of the invention is more beneficial in clinical use compared to bispecific antibodies and known trispecific antibodies, makes easier clearing and expression of an antibody, which can further be used in treating CEA-mediated tumours.
10 cl, 21 dwg, 11 ex
SUBSTANCE: strain A-4A7 is prepared by fusion of mouse myeloma cells of line SP2/0.Agl4 with mouse lymphocytes of line Balb/c immunised by introduction in pads of a purified preparation AMGF (alpha2-microglobulin of fertility) separated from amniotic fluid, and deposited in the transplantable mammal cell culture collection of the Research Institute of Human morphology of the Russian Academy of Medical Science numbered 131/2002. Strain A-4A7 synthesises monoclonal antibodies (MCA) of IgGI class specifically reacting in solid-phase immune-enzyme analysis (IEA) with AMGF isoforms of endometrial, follicular and sperm nature. Activity of the strain: cultural supernatant contains MCA 3-5 mkg/ml, while ascetic fluid contains MCA 2-5 mg/ml. The antibody titre in cultural fluid is 1:500-1:1000, in ascetic fluid up to 1:1×107. A-4A7 bonds various AMGF protein glycoforms produced in male and female reproductive organs. Application of MCA A-4A7 as an immunodiagnosis test systems allows for high-specific and high-sensitive (1 ng/ml) quantitative analysis of various AMGF/glykodeline isoforms in biological liquids.
EFFECT: new compounds are characterised with valuable biological properties.
SUBSTANCE: method for making polyclonal Nogo protein antibodies by protein immunisation of an animal, with at least 6 amino-acid residues of active area Nogo A, free from any other myelin material of central nervous system whereto bound in natural conditions. Said active area covers position 1-171, 260-974, 1163-1178 of the corresponding amino acid sequence of protein Nogo A. There is disclosed separated antiserum based on polyclonal antibodies made under said method. There are disclosed methods for immunisation of a non-human animal to make polyclonal antibodies, as well as method for making monoclonal antibody, and the specified monoclonal antibody.
EFFECT: making active protein Nogo A antibodies to be applied in medicine for neurite growth activation.
26 cl, 18 dwg, 2 tbl, 8 ex
SUBSTANCE: polypeptides include single-domain antibody against vWF, A1 domain of vWF, A1 domain of activated vWF, A3 domain of vWF, gp1b and/or collagen. Invention claims methods of obtaining indicated polypeptides, methods of coating devices applied in medical practice (e.g. in X-ray structural analysis, endoprosthetics) with indicated polypeptides.
EFFECT: obtainment of polypeptides for treatment of diseases requiring modulation of thrombocyte-mediated aggregation.
40 cl, 69 ex, 30 dwg, 32 tbl
SUBSTANCE: claimed invention relates to field of biotechnology and immunology. Described is physiologically active protein conjugate. Protein conjugate includes physiologically active polypeptide, which is covalently connected with Fc fragment of immunoglobulin by means of polyethylene glycol. Described is method of obtaining protein conjugate. It can find application in production of various polypeptide medications of prolonged action.
EFFECT: increased physiological activity in vivo construction in comparison with native physiologically active polypeptide and increase of half-life in serum of physically active polypeptide with minimal risk of inducing undesirable immune response.
17 cl, 18 dwg, 8 ex
FIELD: biotechnology, medicine, proteins.
SUBSTANCE: invention describes new polypeptide in isolated form relating to subfamily of superfamily human immunoglobulins (Ig-Sf). This polypeptide shows at least 70% of homology level with amino acid sequence of murine molecules CRAM-1 or CRAM-2 regulated by the confluence of adhesive (figures 3, 6 are represented in the claim). Also, invention relates to antibodies showing specificity with respect to the polypeptide. Antibodies and soluble polypeptide can be used for treatment of inflammation and tumors. Invention describes polynucleotide or oligonucleotide encoding the full-size polypeptide or its moiety and represents primer, probe, anti-sense RNA and shows the nucleotide sequence that is identical conceptually with human CRAM-1. Invention provides preparing new adhesive proteins from superfamily Ig-Sf that are regulated at the transcription level in endothelium by effect of tumors. Invention can be used for treatment of different diseases, in particular, inflammatory responses.
EFFECT: valuable medicinal properties of polypeptide.
19 cl, 33 dwg, 1 ex
FIELD: immunology; treatment of mediated diseases IL-1 and failures.
SUBSTANCE: bonding molecule IL-1β which is antibody to human IL-1β and especially human antibody to human IL-1β where hypervariable sections CDRs of heavy and light chains have definite amino acid sequences. Antibody may be used for treatment of mediated disease IL-1, for example osteoarthritis, osteoporosis and other inflammatory processes of bones of rheumatism or podagra nature. Constructions of deoxyribonucleic acid are described which code heavy and light chains or their fragments and expressive vectors which may be replicated in cells including deoxyribonucleic acid constructions. Method of obtaining bonding molecule IL-1β by means of cell transformed by vector is described. Proposed antibody may be used both in prophylactic and treatment of diseases.
EFFECT: enhanced efficiency.
15 cl, 3 dwg, 5 ex
FIELD: medicine, oncology, gastroenterology, immunobiotechnology.
SUBSTANCE: invention describes an antibody or its derivative, or its fragment showing the structure able to bind the target structure. Antibody is located inside and on surface of human gastroenteric tract epithelial tumor cells and in subpopulation of normal gastroenteric tract epithelial cells. Indicated binding structures comprise sequences determining the complementarity of the region (CDR) in light chain comprising in main amino acids at number 23-33 (CDR 1), 49-55 (CDR 2), 88-98 (CDR 3) of amino acid sequence represented in SEQ ID NO:2, and CDR sequence in heavy chains comprising in main amino acids at number 158-162 (CDR 1), 177-193 (CDR 2), 226-238 (CDR 3) of amino acid sequence represented in SEQ ID NO:2, or other binding structures with similar unique binding properties. Also, invention describes the target-structure located inside or on surface of tumor cells: vaccine composition designated for treatment of malignant disease in human and comprising abovementioned antibody. Also, invention describes methods for treatment and diagnosis of malignant disease. Using this invention provides preparing antibodies that relieve identification of new phenotype-specific tumor-associated antigens, to predict and treat metastatic human diseases. Invention can be used in medicinal practice.
EFFECT: valuable medicinal properties of antibodies.
37 cl, 21 dwg, 4 tbl, 8 ex
FIELD: medicine, oncology, immunology.
SUBSTANCE: invention relates to humanized antibodies with ErbB2. Invention involves the development of new humanized antibodies raised to tyrosinase receptors of family ErbB2, and to a composition comprising these antibodies. The advantage of invention involves expanding region in using indicated antibodies in cancer treatment wherein receptor of epidermal growth factor, EGFR, is a target of these antibodies.
EFFECT: valuable properties of antibody.
14 cl, 3 tbl, 13 dwg
FIELD: biology, medicine, immunology.
SUBSTANCE: invention relates to a method for preparing antiserum against mammal CNSS peptides. Method is carried out by immunization of mammals (man, cat, pig) with CNSS peptides in combination with full Freund's adjuvant by subcutaneous and subconjunctive administration and by intravenous administration as a pure form. Antigen is administrated to the following mammals: rabbit, guinea pig and ass for three steps with break for 2 days between each step. Two days after the last administration of antigen antiserum against vertebrate CNSS peptides with a titer value 1:256 is prepared from heart of live fixed animal and stored it in native or lyophilized state at 4-5°C. Method provides preparing antiserum with high content of specific antibodies for short time, method shows low difficulty and economy and sterile serum can be stored for up to 2-3 years without loss of its activity.
EFFECT: improved preparing method.
FIELD: biotechnology, molecular biology, medicine.
SUBSTANCE: invention discloses amino acid sequences of human obesity polypeptide (OB) two isoforms possessing capacity for modulation of animal body mass, their signal peptide-containing precursors and analogues. Polypeptide isoforms are prepared as result of insertions, deletions and amino acid changes that retain activity typical for nonmodified forms of OB-polypeptides, and polyclonal and monoclonal antibodies interaction specifically with new agents modulating the body mass value also. Invention describes DNA sequences encoding these polypeptides and their analogues, vector structures comprising these sequences used for preparing recombinant forms of OB-polypeptides. Invention proposes using new polypeptides and their analogues as an active component in pharmaceutical compositions. Using this invention can promote to solving the problem for providing medicine, veterinary science and animal husbandry with effective agent used for decreasing the body mass value. Invention can be used in medicine for diagnosis and treatment of pathological states associated with disturbance of regulation of human body mass, and in animal husbandry and veterinary science.
EFFECT: valuable biological, medicinal and veterinary properties of polypeptide.
23 cl, 71 dwg, 12 tbl, 17 ex
SUBSTANCE: invention relates to proteins and polynucleotides, which stimulate enzyme cleavage and releasing of TNF receptors. Also disclosed are methods for identification of additional agents, which influence on TNF receptor releasing from cells. Products of present invention containing in pharmaceutical compositions as active ingredients increase or decrease TNF signal transduction and consequently abate disease pathology.
EFFECT: new pharmaceutical compositions.
27 cl, 8 ex, 3 tbl, 5 dwg
FIELD: medicine, biotechnology.
SUBSTANCE: invention relates to antibodies specifically binding to new human extracellular matrix polypeptides called as RGI; immunoconjugate containing the same and method for selective cell degradation; method for treatment of prostates cancer and metastasis in patients suffering from prostates cancer.
EFFECT: new method for treatment of prostates cancer.
28 cl, 7 ex, 7 dwg
FIELD: biotechnology, medicine, immunology.
SUBSTANCE: invention relates to a modified antibody comprising two or more V-regions in H-chain and two or more V-regions in L-chain of antibody bound directly or over a linker by covalent or noncovalent bond. Antibody has a lesser size as compared with the parent antibody possessing or not possessing the agonistic effect to TPO receptors. Antibody represents TPO agonist and able for specific recognizing and cross-linking the TPO receptor. For induction of the agonistic effect in cells expressing TPO receptors cells are contacted with the modified antibody. Measurement of the antibody TRO-agonistic effect is carried out by cross-linking TRO receptors. The modified antibody can be used as a medicinal agent in treatment of thrombocytopenia. Modified antibodies possess higher activity as compared whole antibodies (JgG) and improved penetration capacity into tissues owing to decreased molecular sizes and absence of constant regions.
EFFECT: improved and valuable properties of antibody.
38 cl, 92 dwg, 3 tbl, 8 ex
FIELD: biochemistry, biotechnology, peptides.
SUBSTANCE: invention relates to protein of molecular mass 12000 Da isolated from medicinal leech saliva (Hirudo medicinalis) comprising 6 cysteine residues able to form bonds -S-S-, and pI value about 3.7. The novel protein possesses ability to block platelets adhesion induced by collagen. As result of screening cDNA library of H. medicinalis a novel gene of new inhibitor of adhesion is identified and its nucleotide sequence is determined. Invention describes a method for preparing recombinant form of protein involving transformation of suitable cells with vector comprising DNA sequence encoding inhibitor, culturing cells under condition providing its expression and isolation of expressed protein. Invention proposes using natural and recombinant forms of inhibitor in therapy states associated with vessels congestion and diseases of circulatory system, and for treatment of articles surface made of natural and artificial collagen.
EFFECT: valuable biological and medicinal properties of polypeptide.
17 cl, 10 dwg, 13 ex