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Epitope mapping

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Abstract

This article describes a strategy for the mapping of the binding site, or epitope, of a monoclonal antibody (MAb) using bacterially expressed protein products. An overall strategy is discussed. This includes an initial round of several parallel approaches to gain the greatest amount of information at this stage. The second round uses the mapping information generated to identify MAbs, which may bind to identical or overlapping epitopes. The third round involves the design of new constructs that express small defined regions of the protein to refine the position of the epitope. The final step leads to the identification of the epitope to a resolution of 10 amino acid residues or else.

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References

  1. Ruther, U. and Miiller-Hill, B. (1983) Easy identification of cDNA clones.EMBO J. 2, 1791–1794.

    CAS  Google Scholar 

  2. Yanisch-Perron, C., Vieira, J., and Messing, J. (1985) Improved M13 phage cloning vectors and host strains: Nucleotide sequences of the M13mp l8 and pUC19 vectors.Gene 33, 103–119.

    Article  CAS  Google Scholar 

  3. Tabor, S. and Richardson, C. C. (1985) A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes.Proc. Natl. Acad. Sci. 82, 1074–1078.

    Article  CAS  Google Scholar 

  4. Stratagene catalogue (1989), pp. 156-159.

  5. Cunningham, B. C., Jhurani, P., Ng, P., and Wells, J. A. (1989) Receptor and antibody epitopes in human growth hormone identified by homolog-scanning mutagenesis.Science 243, 1330–1336.

    Article  CAS  Google Scholar 

  6. Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989)Molecular Cloning: A Laboratory Manual, 2nd Ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.

    Google Scholar 

  7. Harlow, E. and Lane, D. P. (1989)Antibodies: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

    Google Scholar 

  8. Mole, S. E, Iggo, R. D, and Lane D. P. (1989) Using the polymerase chain reaction to modify expression plasmids for epitope mapping.Nucleic Acids Res. 17, 3319.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Pediatrics, University College London Medical School, The Rayne Institute, University Street, WC1E 6JJ, London, UK

    Sara E. Mole

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  1. Sara E. Mole
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Mole, S.E. Epitope mapping. Mol Biotechnol 1, 277–287 (1994). https://doi.org/10.1007/BF02921695

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  • DOI: https://doi.org/10.1007/BF02921695

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