Antibodies and kin molecules bound to psca proteins

FIELD: biotechnologies.

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.



 

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