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).
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
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.
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.
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.
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.
Kyte, J. and Doolittle, R.F. (1982) A simple method for displaying the hydropathic character of a protein. J. Mol. Biol. 157, 105–132.
Hopp, T.P. (1986) Protein surface analysis. J. Immunol. Meth. 88, 1–18.
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.
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.
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.
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.
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.
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.
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.
Merrifield, B. (1986) Solid phase synthesis. Science 232, 341–347.
Atherton, E. and Sheppard, R.C. (1985) Solid phase peptide synthesis using N-fluorenylmethoxycarbonylamino acid pentaflurophenyl esters. J. Chem. Soc. Chetn. Commun. 165–166.
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.
Kagan, A. and Glick, M. (1974) Oxytocin, in Methods in Hormone Radioimmunoassay (Jaffe, B.M. and Behrman, H. R., eds.) Academic, New York.
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.
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.
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.
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.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1988 The Humana Press Inc.
About this protocol
Cite this protocol
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
Download citation
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