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. 1993 Mar;78(3):364–370.

Construction of a reshaped HMFG1 antibody and comparison of its fine specificity with that of the parent mouse antibody.

M E Verhoeyen 1, J A Saunders 1, M R Price 1, J D Marugg 1, S Briggs 1, E L Broderick 1, S J Eida 1, A T Mooren 1, R A Badley 1
PMCID: PMC1421827  PMID: 7682986

Abstract

A human antibody with milk mucin specificity was obtained by transferring the complementarity determining regions (CDR) of the mouse antibody HMFG1 onto carefully selected human framework regions. The resulting reshaped human antibody, HuHMFG1, showed no difference in relative affinity for its antigen compared with the parent mouse HMFG1. The minimum epitope recognized by both the mouse and reshaped antibodies was demonstrated by epitope mapping to be identical, and consists of the tetramer PDTR. In a replacement net analysis, in which each of the amino acids was replaced in turn with the 19 other residues, it was determined that mouse HMFG1 and HuHMFG1 reacted with this series of synthetic peptides in an equivalent manner, indicating retention of identical fine specificity in the HuHMFG1 antibody. In contrast to other published reports, this was achieved without involvement of any framework residues in the binding site transfer. These data demonstrate that if well-matching human framework regions are employed grafting the CDR only can be sufficient to confer desired specificities to human antibodies and can, indeed, provide human analogues of mouse antibodies with virtually indistinguishable affinities and fine specificities relative to the mouse parent antibodies.

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Selected References

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

  1. Briggs S., Price M. R., Tendler S. J. Immune recognition of linear epitopes in peptide fragments of epithelial mucins. Immunology. 1991 Aug;73(4):505–507. [PMC free article] [PubMed] [Google Scholar]
  2. Burchell J., Gendler S., Taylor-Papadimitriou J., Girling A., Lewis A., Millis R., Lamport D. Development and characterization of breast cancer reactive monoclonal antibodies directed to the core protein of the human milk mucin. Cancer Res. 1987 Oct 15;47(20):5476–5482. [PubMed] [Google Scholar]
  3. Carter P., Presta L., Gorman C. M., Ridgway J. B., Henner D., Wong W. L., Rowland A. M., Kotts C., Carver M. E., Shepard H. M. Humanization of an anti-p185HER2 antibody for human cancer therapy. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4285–4289. doi: 10.1073/pnas.89.10.4285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Co M. S., Avdalovic N. M., Caron P. C., Avdalovic M. V., Scheinberg D. A., Queen C. Chimeric and humanized antibodies with specificity for the CD33 antigen. J Immunol. 1992 Feb 15;148(4):1149–1154. [PubMed] [Google Scholar]
  5. Co M. S., Deschamps M., Whitley R. J., Queen C. Humanized antibodies for antiviral therapy. Proc Natl Acad Sci U S A. 1991 Apr 1;88(7):2869–2873. doi: 10.1073/pnas.88.7.2869. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Epp O., Colman P., Fehlhammer H., Bode W., Schiffer M., Huber R., Palm W. Crystal and molecular structure of a dimer composed of the variable portions of the Bence-Jones protein REI. Eur J Biochem. 1974 Jun 15;45(2):513–524. doi: 10.1111/j.1432-1033.1974.tb03576.x. [DOI] [PubMed] [Google Scholar]
  7. Gendler S., Taylor-Papadimitriou J., Duhig T., Rothbard J., Burchell J. A highly immunogenic region of a human polymorphic epithelial mucin expressed by carcinomas is made up of tandem repeats. J Biol Chem. 1988 Sep 15;263(26):12820–12823. [PubMed] [Google Scholar]
  8. Geysen H. M., Rodda S. J., Mason T. J., Tribbick G., Schoofs P. G. Strategies for epitope analysis using peptide synthesis. J Immunol Methods. 1987 Sep 24;102(2):259–274. doi: 10.1016/0022-1759(87)90085-8. [DOI] [PubMed] [Google Scholar]
  9. Gorman S. D., Clark M. R., Routledge E. G., Cobbold S. P., Waldmann H. Reshaping a therapeutic CD4 antibody. Proc Natl Acad Sci U S A. 1991 May 15;88(10):4181–4185. doi: 10.1073/pnas.88.10.4181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  11. Hieter P. A., Max E. E., Seidman J. G., Maizel J. V., Jr, Leder P. Cloned human and mouse kappa immunoglobulin constant and J region genes conserve homology in functional segments. Cell. 1980 Nov;22(1 Pt 1):197–207. doi: 10.1016/0092-8674(80)90168-3. [DOI] [PubMed] [Google Scholar]
  12. Hird V., Verhoeyen M., Badley R. A., Price D., Snook D., Kosmas C., Gooden C., Bamias A., Meares C., Lavender J. P. Tumour localisation with a radioactively labelled reshaped human monoclonal antibody. Br J Cancer. 1991 Nov;64(5):911–914. doi: 10.1038/bjc.1991.425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jones P. T., Dear P. H., Foote J., Neuberger M. S., Winter G. Replacing the complementarity-determining regions in a human antibody with those from a mouse. 1986 May 29-Jun 4Nature. 321(6069):522–525. doi: 10.1038/321522a0. [DOI] [PubMed] [Google Scholar]
  14. Kettleborough C. A., Saldanha J., Heath V. J., Morrison C. J., Bendig M. M. Humanization of a mouse monoclonal antibody by CDR-grafting: the importance of framework residues on loop conformation. Protein Eng. 1991 Oct;4(7):773–783. doi: 10.1093/protein/4.7.773. [DOI] [PubMed] [Google Scholar]
  15. Maeda H., Matsushita S., Eda Y., Kimachi K., Tokiyoshi S., Bendig M. M. Construction of reshaped human antibodies with HIV-neutralizing activity. Hum Antibodies Hybridomas. 1991 Jul;2(3):124–134. [PubMed] [Google Scholar]
  16. Price M. R., Hudecz F., O'Sullivan C., Baldwin R. W., Edwards P. M., Tendler S. J. Immunological and structural features of the protein core of human polymorphic epithelial mucin. Mol Immunol. 1990 Aug;27(8):795–802. doi: 10.1016/0161-5890(90)90089-i. [DOI] [PubMed] [Google Scholar]
  17. Price M. R., Sekowski M., Tendler S. J. Purification of anti-epithelial mucin monoclonal antibodies by epitope affinity chromatography. J Immunol Methods. 1991 May 17;139(1):83–90. doi: 10.1016/0022-1759(91)90354-i. [DOI] [PubMed] [Google Scholar]
  18. Queen C., Schneider W. P., Selick H. E., Payne P. W., Landolfi N. F., Duncan J. F., Avdalovic N. M., Levitt M., Junghans R. P., Waldmann T. A. A humanized antibody that binds to the interleukin 2 receptor. Proc Natl Acad Sci U S A. 1989 Dec;86(24):10029–10033. doi: 10.1073/pnas.86.24.10029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Riechmann L., Clark M., Waldmann H., Winter G. Reshaping human antibodies for therapy. Nature. 1988 Mar 24;332(6162):323–327. doi: 10.1038/332323a0. [DOI] [PubMed] [Google Scholar]
  20. Routledge E. G., Lloyd I., Gorman S. D., Clark M., Waldmann H. A humanized monovalent CD3 antibody which can activate homologous complement. Eur J Immunol. 1991 Nov;21(11):2717–2725. doi: 10.1002/eji.1830211111. [DOI] [PubMed] [Google Scholar]
  21. Saul F. A., Amzel L. M., Poljak R. J. Preliminary refinement and structural analysis of the Fab fragment from human immunoglobulin new at 2.0 A resolution. J Biol Chem. 1978 Jan 25;253(2):585–597. [PubMed] [Google Scholar]
  22. Scanlon M. J., Morley S. D., Jackson D. E., Price M. R., Tendler S. J. Structural and computational investigations of the conformation of antigenic peptide fragments of human polymorphic epithelial mucin. Biochem J. 1992 May 15;284(Pt 1):137–144. doi: 10.1042/bj2840137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shawler D. L., Bartholomew R. M., Smith L. M., Dillman R. O. Human immune response to multiple injections of murine monoclonal IgG. J Immunol. 1985 Aug;135(2):1530–1535. [PubMed] [Google Scholar]
  24. Shearman C. W., Pollock D., White G., Hehir K., Moore G. P., Kanzy E. J., Kurrle R. Construction, expression and characterization of humanized antibodies directed against the human alpha/beta T cell receptor. J Immunol. 1991 Dec 15;147(12):4366–4373. [PubMed] [Google Scholar]
  25. Takahashi N., Ueda S., Obata M., Nikaido T., Nakai S., Honjo T. Structure of human immunoglobulin gamma genes: implications for evolution of a gene family. Cell. 1982 Jun;29(2):671–679. doi: 10.1016/0092-8674(82)90183-0. [DOI] [PubMed] [Google Scholar]
  26. Taylor-Papadimitriou J., Peterson J. A., Arklie J., Burchell J., Ceriani R. L., Bodmer W. F. Monoclonal antibodies to epithelium-specific components of the human milk fat globule membrane: production and reaction with cells in culture. Int J Cancer. 1981 Jul 15;28(1):17–21. doi: 10.1002/ijc.2910280104. [DOI] [PubMed] [Google Scholar]
  27. Tempest P. R., Bremner P., Lambert M., Taylor G., Furze J. M., Carr F. J., Harris W. J. Reshaping a human monoclonal antibody to inhibit human respiratory syncytial virus infection in vivo. Biotechnology (N Y) 1991 Mar;9(3):266–271. doi: 10.1038/nbt0391-266. [DOI] [PubMed] [Google Scholar]
  28. Verhoeyen M., Milstein C., Winter G. Reshaping human antibodies: grafting an antilysozyme activity. Science. 1988 Mar 25;239(4847):1534–1536. doi: 10.1126/science.2451287. [DOI] [PubMed] [Google Scholar]

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