Investigating the Specificity of Monoclonal Antibodies to Protein Antigens Using β-Galactosidase Fusion Proteins

  • D. P. Lane
  • J. Gannon
  • S. P. J. Brennan
  • S. E. Mole
Part of the Methodological Surveys in Biochemistry and Analysis book series (MSBA, volume 15B)

Abstract

Numerous monoclonal antibodies (MAb’s) to the oncogene, large T, of the small double-stranded DNA (dsDNA) tumour virus SV40 have been produced [1–4, and J. Yewdell & D.P.Lane, unpublished work). Detailed analysis of these Ab’s* has provided insights into both the structure and the function of large T, besides protein-antigen immunochemistry. The Ab’s can be subdivided by the following criteria.— (a) Ability of the Ab to recognize the protein after denaturation. (b) Recognition of only a sub-set of the T antigen molecules. (c) Inhibition of biological activity of large T. (d) Location of the binding site of the Ab on the linear structure and the 3-dimensional structure of the protein. Each subdivision can be enhanced by subcloning fragments of the large-T coding information into bacterial and eukaryotic expression vectors that cause the large-T fragment to be incorporated at the carboxy terminus of β-galactosidase [5, 6].

Keywords

Amide Influenza Electrophoresis Paration Alan 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Lane, D.P. & Hoeffler, W.K. (1980) Nature 288, 167–170.CrossRefGoogle Scholar
  2. 2.
    Gurney, E.G., Harrison, R.O. & Fenno, J. (1980) J. Virol. 34, 752–763.Google Scholar
  3. 3.
    Harlow, E., Crawford, L.V., Pim, D.C. & Williamson, N.M. (1981) J. Virol. 39, 861–869.Google Scholar
  4. 4.
    Ball, R.K., Siegl, R., Quellhorst, S., Branpner, G. & Braun, D.G. (1984) EMBO J. 3, 1485–1491.Google Scholar
  5. 5.
    Rüther, U. & Müller-Hill, B. (1983) EMBO J. 2, 1791–1794.Google Scholar
  6. 6.
    Mole, S.E. & Lane, D.P. (1985) J. Virol. 52,in press.Google Scholar
  7. 7.
    Towbin, H., Staehlin, T. & Gordon, J. (1979) Proc. Nat. Acad. Sci. 76, 4350–4354.CrossRefGoogle Scholar
  8. 8.
    Lane, D.P. & Robbins, A.K. (1978) Virology 87, 182–193.CrossRefGoogle Scholar
  9. 9.
    Rigby, P.W.J. & Lane, D.P. (1983) in Advances in Viral Oncology, Vol. 3 (Klein, G., ed.), Raven Press, New York, pp. 31–57.Google Scholar
  10. 10.
    Benchimol, S., Pim, D. & Crawford, L.V. (1982) EMBO J. 1, 1052–1062.Google Scholar
  11. 11.
    Dippold, W.G., Jay, G., De Leo, A.B., Khoury, G. & Old, L.J. (1981) Proc. Nat. Acad. Sci. 78, 1695–1699.CrossRefGoogle Scholar
  12. 12.
    Dilworth, S.M. & Griffen, B.E. (1982) Proc. Nat. Acad. Sci. 79, 1059–1062.CrossRefGoogle Scholar
  13. 13.
    Thomas, R., Kaplan, L., Reich, N., Lane, D.P. & Levine, A.J. (1983) Virology 131, 502–517.CrossRefGoogle Scholar
  14. 14.
    Holmes, N.J. & Parham, P. (1983) J. Biol. Chem. 256, 1580–1586.Google Scholar
  15. 15.
    Thomson, R.J. & Jackson, A.P. (1984) Trends in Biochem. Sci. 9, 1–3.CrossRefGoogle Scholar
  16. 16.
    Clark, R., Lane, D.P. & Tjian, R. (1981) J. Biol. Chem. 256,11854–11858.Google Scholar
  17. 17.
    Crawford, L., Leppard, K., Lane, D. & Harlow, E. (1983) in Tumor viruses and Differentiation (Scolnick, E.M. & Levine, A.J., eds.), Alan Liss, New York, pp. 421–425.Google Scholar
  18. 18.
    Clark, R., Peden, K., Pipas; J.M., Nathans, D. & Tjian, R. (1983) Mol. Cell. Biol. 3, 220–228.Google Scholar
  19. 19.
    Stanley, K.K. (1983) Nucleic Acids Res. 11, 4077–4092.CrossRefGoogle Scholar
  20. 20.
    Nunberg, J.H., Rodgers, G., Gilbert, J.H. & Snead, R.M. (1984) Proc. Nat. Acad. Sci. 81, 3675–3679.CrossRefGoogle Scholar
  21. 21.
    Hall, C.V., Jacob, P.E., Ringold, G.M. & Lee, F. (1983) J. Mol. Appl. Genet. 2, 101–109.Google Scholar
  22. 22.
    Wilson, I.A., Niman, H.L., Houghten, R.A., Cherenson, A.R., Connnolly, M.L. & Lerner, R.A. (1984) Cell 37, 767–778.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1985

Authors and Affiliations

  • D. P. Lane
    • 1
  • J. Gannon
    • 1
  • S. P. J. Brennan
    • 1
  • S. E. Mole
    • 1
  1. 1.Department of Biochemistry Cancer Research Campaign Eukaryotic Molecular Genetics Research GroupImperial CollegeLondonUK

Personalised recommendations