Advertisement

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large T, of the small double-stranded DNA (dsDNA) tumour virus SV40 have been produced [1–4, and J. Yewdell & D.P.Lane, unpublished work).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

Fusion Protein Protein Antigen Carboxy Terminus Eukaryotic Expression Vector Cancer Research Campaign 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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