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Production of Antisera to Synthetic Peptides

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 3))

Abstract

The objective of immunization is to produce polyclonal or monoclonal antibodies that react with a chosen molecule. Immunization may be in vivo by injecting antigen into animals to produce antisera or to provide B-lymphocytes for immortalization as hybridomas, or alternatively, in vitro using splenocytes from untreated animals (1) for subsequent fusion with myeloma cells to produce monoclonal antibodies (MAbs).

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References

  1. Luben, R.A., Brazeau, P. Bolen, P., and Guillemin, R. (1982) Monoclonal antibodies to hypothalamic growth hormone releasing factor with picomoles of antigen. Science 218, 887–889.

    Article  PubMed  CAS  Google Scholar 

  2. Tzartos, S.J. and Lindstrom, J.M. (1980) Monoclonal antibodies used to probe acetylcholine receptor structure: localization of the main immunogenic region and detection of similarities between subunits. Proc. Natl. Acad. Sci. USA 77, 755–759.

    Article  PubMed  CAS  Google Scholar 

  3. Gullick, W.J. and Lindstrom, J.M. (1983) Mapping the binding of monoclonal antibodies to the acetylcholine receptor from Torpedo californica. Biochemistry 22, 3312–3320.

    Article  PubMed  CAS  Google Scholar 

  4. Hopp, T.P. and Woods, K.R. (1981) Prediction of protein antigenic determinants from amino acid sequences. Proc. Natl. Acad. Sci. USA 78, 3824–3828.

    Article  PubMed  CAS  Google Scholar 

  5. Kyte, J. and Doolittle, R.F. (1982) A simple method for displaying the hydropathic character of a protein. J. Mol. Biol. 157, 105–132.

    Article  PubMed  CAS  Google Scholar 

  6. Hopp, T.P. (1986) Protein surface analysis. J. Immunol. Meth. 88, 1–18.

    Article  CAS  Google Scholar 

  7. Thornton, J.M., Edwards, M.S., Taylor, W.R., and Barlow, DJ. (1986) Location of “continuous” antigenic determinants in the protruding regions of proteins. EMBO J. 5, 409–413.

    PubMed  CAS  Google Scholar 

  8. Schulze-Gahmen, U., Prinz, H., Glatter, U., and Beyreuther, K. (1985) Towards assignment of secondary structures by anti-peptide antibodies. Specificity of the immune response to a turn. EMBO J. 4, 1731–1737.

    PubMed  CAS  Google Scholar 

  9. Westhof, E., Altschuh, D., Moras, D., Bloomer, A.C., Mon-dragon, A., Klug, A., and Van Regenmortel, M.H.V. (1984) Correlation between segmental mobility and the location of antigenic determinants in proteins. Nature 311, 123–126.

    Article  PubMed  CAS  Google Scholar 

  10. Tainer, J.A., Getzoff, E.D., Alexander, H., Houghton, R.A., Olson, A.J., Lerner, R.A., and Hendrickson, W.A. (1984) The reactivity of anti-peptide antibodies is a function of the atomic mobility of sites in a protein. Nature 312, 127–134.

    Article  PubMed  CAS  Google Scholar 

  11. Sternberg, M.J.E. and Taylor, W.R. (1984) Modelling the ATP-binding site of oncogene products, the epidermal growth factor receptor and related proteins. FEBS Lett. 175, 387–392.

    Article  PubMed  CAS  Google Scholar 

  12. Gullick, W.J., Downward, J., Foulkes, J.G., and Waterfield, M.D. (1986) Antibodies to the ATP-binding site of human epidermal growth factor receptor as specific inhibitors of EGF stimulated protein tyrosine kinase activity. Eur. J. Biochetn. 158, 245–253.

    Article  CAS  Google Scholar 

  13. Gullick, W.J., Downward, J., and Waterfield, M.D. (1985) Antibodies to the autophosphorylation sites of the epidermal growth factor receptor as probes of structure and function. EMBO J. 4, 2869–2877.

    PubMed  CAS  Google Scholar 

  14. Merrifield, B. (1986) Solid phase synthesis. Science 232, 341–347.

    Article  PubMed  CAS  Google Scholar 

  15. Atherton, E. and Sheppard, R.C. (1985) Solid phase peptide synthesis using N-fluorenylmethoxycarbonylamino acid pentaflurophenyl esters. J. Chem. Soc. Chetn. Commun. 165–166.

    Google Scholar 

  16. Palfreyman, J.W., Aitcheson, T.C., and Taylor, P. (1984) Guidelines for the production of polypeptide specific antisera using small synthetic oligopeptides as immunogens. J. Immunol. Meth. 75, 383–393.

    Article  CAS  Google Scholar 

  17. Kagan, A. and Glick, M. (1974) Oxytocin, in Methods in Hormone Radioimmunoassay (Jaffe, B.M. and Behrman, H. R., eds.) Academic, New York.

    Google Scholar 

  18. Kris, R.M., Lax, I., Sasson, I., Copf, B., Welin, S., Gullick, W.J., Waterfield, M.D., Ullrich, A., Fridkin, M., and Schlessinger, J. (1985) Synthetic peptide approach to the analysis of the kinase activities of avian EGF receptor and v-erbB protein. Biochemie 67, 1095–1101.

    Article  CAS  Google Scholar 

  19. Green, N., Alexander, H., Olson, A., Alexander, S., Shinnick, T.M., Sutcliffe, J.G., and Lerner, R.A. (1982) Immunogenic structure of the Influenza virus hemagglutinin. Cell 28, 477–487.

    Article  PubMed  CAS  Google Scholar 

  20. Ratam, M. and Lindstrom, J. (1984) Structural features of the nicotinic acetylcholine receptor revealed by antibodies to synthetic peptides. Biochem. Biophys. Res. Comm. 122, 1225–1233.

    Article  Google Scholar 

  21. Gullick, W.J., Marsden J.J., Whittle, N., Ward, B., Bobrow, L., and Waterfield, M.D. (1986) Expression of epidermal growth factor receptors on cervical, ovarian and vulval carcinomas. Cancer Res. 46, 285–292.

    PubMed  CAS  Google Scholar 

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© 1988 The Humana Press Inc.

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Gullick, W.J. (1988). Production of Antisera to Synthetic Peptides. In: Walker, J.M. (eds) New Protein Techniques. Methods in Molecular Biology™, vol 3. Humana Press. https://doi.org/10.1385/0-89603-126-8:341

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  • DOI: https://doi.org/10.1385/0-89603-126-8:341

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-126-5

  • Online ISBN: 978-1-59259-490-0

  • eBook Packages: Springer Protocols

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