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. 1990 Apr;61(4):508–513. doi: 10.1038/bjc.1990.115

A bispecific monoclonal antibody against methotrexate and a human tumour associated antigen augments cytotoxicity of methotrexate-carrier conjugate.

M V Pimm 1, R A Robins 1, M J Embleton 1, E Jacobs 1, A J Markham 1, A Charleston 1, R W Baldwin 1
PMCID: PMC1971384  PMID: 2331436

Abstract

A bispecific monoclonal antibody, reactive with methotrexate (MTX) and a tumour associated antigen (gp72) has been produced by fusing spleen cells from MTX immunised mice with 791T/36/3 (anti-gp72) hybridoma. The hybrid antibody was purified from anti-MTX and anti-gp72 antibodies present in the hybridoma culture supernatant by combinations of affinity chromatography on a MTX-agarose immunoabsorbent and stepwise acid elution from Sepharose-Protein A. A particular feature of the present antibody is that it reacts with conjugated MTX; this would allow in vivo targeting of conjugates, increasing many fold the number of molecules of drug carried or localising to pre-targeted antibody. Dual binding between tumour cell surface antigen and MTX was demonstrated by the ability of the hybrid antibody to bridge between tumour cells and MTX as MTX-HSA conjugate, reaction here being detected by flow cytofluorimetry. Purified hybrid antibody specifically enhanced the in vitro cytotoxicity of MTX-HSA for gp72 positive tumour cells.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baldwin R. W., Byers V. S., Pimm M. V. Monoclonal antibodies and immunoconjugates for cancer treatment. Cancer Chemother Biol Response Modif. 1988;10:397–415. [PubMed] [Google Scholar]
  2. Ballantyne K. C., Perkins A. C., Pimm M. V., Garnett M. C., Clegg J. A., Armitage N. C., Baldwin R. W., Hardcastle J. D. Biodistribution of a monoclonal antibody-methotrexate conjugate (791T/36-MTX) in patients with colorectal cancer. Int J Cancer Suppl. 1988;2:103–108. doi: 10.1002/ijc.2910410724. [DOI] [PubMed] [Google Scholar]
  3. Corvalan J. R., Smith W. Construction and characterisation of a hybrid-hybrid monoclonal antibody recognising both carcinoembryonic antigen (CEA) and vinca alkaloids. Cancer Immunol Immunother. 1987;24(2):127–132. doi: 10.1007/BF00205589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Corvalan J. R., Smith W., Gore V. A., Brandon D. R., Ryde P. J. Increased therapeutic effect of vinca alkaloids targeted to tumour by a hybrid-hybrid monoclonal antibody. Cancer Immunol Immunother. 1987;24(2):138–143. doi: 10.1007/BF00205591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Corvalan J. R., Smith W., Gore V. A., Brandon D. R. Specific in vitro and in vivo drug localisation to tumour cells using a hybrid-hybrid monoclonal antibody recognising both carcinoembryonic antigen (CEA) and vinca alkaloids. Cancer Immunol Immunother. 1987;24(2):133–137. doi: 10.1007/BF00205590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Embleton M. J., Gunn B., Byers V. S., Baldwin R. W. Antitumour reactions of monoclonal antibody against a human osteogenic-sarcoma cell line. Br J Cancer. 1981 May;43(5):582–587. doi: 10.1038/bjc.1981.87. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Garnett M. C., Embleton M. J., Jacobs E., Baldwin R. W. Preparation and properties of a drug-carrier-antibody conjugate showing selective antibody-directed cytotoxicity in vitro. Int J Cancer. 1983 May 15;31(5):661–670. doi: 10.1002/ijc.2910310520. [DOI] [PubMed] [Google Scholar]
  8. Karawajew L., Micheel B., Behrsing O., Gaestel M. Bispecific antibody-producing hybrid hybridomas selected by a fluorescence activated cell sorter. J Immunol Methods. 1987 Feb 11;96(2):265–270. doi: 10.1016/0022-1759(87)90323-1. [DOI] [PubMed] [Google Scholar]
  9. Pimm M. V., Clegg J. A., Garnett M. C., Baldwin R. W. Biodistribution and tumour localization of a methotrexate-monoclonal-antibody 791T/36 conjugate in nude mice with human tumour xenografts. Int J Cancer. 1988 Jun 15;41(6):886–891. doi: 10.1002/ijc.2910410620. [DOI] [PubMed] [Google Scholar]
  10. Roe R., Robins R. A., Laxton R. R., Baldwin R. W. Kinetics of divalent monoclonal antibody binding to tumour cell surface antigens using flow cytometry: standardization and mathematical analysis. Mol Immunol. 1985 Jan;22(1):11–21. doi: 10.1016/0161-5890(85)90029-x. [DOI] [PubMed] [Google Scholar]
  11. Webb K. S., Ware J. L., Parks S. F., Walther P. J., Paulson D. F. Evidence for a novel hybrid immunotoxin recognizing ricin A-chain by one antigen-combining site and a prostate-restricted antigen by the remaining antigen-combining site: potential for immunotherapy. Cancer Treat Rep. 1985 Jun;69(6):663–672. [PubMed] [Google Scholar]

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