Skip to main content
SpringerLink
Log in

Targeting Antipeptide Antibodies Towards Cytochrome P450 Enzymes

  • Protocol
Cytochrome P450 Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 107))

Abstract

Antibodies against cytochrome P450 enzymes can be produced by immunizing animals with synthetic peptides that mimic small regions of the apoprotein. As the amino-acid sequences of a large number of cytochrome P450 enzymes are known (1), this approach to raising antibodies can be applied to most of the major cytochrome P450 enzymes in human, rat and other species. This is a relattvely simple, rapid and effective method of producmg antibodies.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Nelson, D. R., Koymans, L, Kamataki, T, Stegeman, J. J., Feyeretsen, R, Waxman, D. J., Waterman, M. R, Gotoh, O., Coon, M. J., Estabrook, R. W, Gunsalus, I. C., and Nebert, D. W (1996) P450 superfamily: update on new sequences, gene mapping, accession numbers and nomenclature. Pharmacogenetzcs 6, 1–42

    Article  CAS  Google Scholar 

  2. Parkinson, A. and Gemzlk, B. (1991) Production and purtticatron of anttbodies against rat liver P450 enzymes. Methods Enzymol. 56, 233–245.

    Article  Google Scholar 

  3. Thomas, P. E., Bandiera, S., Reik, L. M., Mames, S. L, Ryan, D. E., and Levm, W. (1987) Polyclonal and monoclonal antibodies as probes of rat hepatic cytochrome P-450 isozymes. Fed Proc. 46, 2563–2566

    PubMed  CAS  Google Scholar 

  4. Wrighton, S. A., Vandenbranden, M., Becker, G, W., Black, S D., and Thomas, P. E. (1992) Two monoclonal antrbodres recognizing different epitopes on rat cytochrome IIB 1 react with human IIE 1. Mol Pharmacol 41, 76–82.

    PubMed  CAS  Google Scholar 

  5. Goldfarb, I., Korzekwa, K., Krausz, K. W, Gonzalez, F., and Gelbom, H V. (1993) Cross-reactivity of thirteen monoclonal anttbodies with ten vaccima cDNA expressed rat, mouse and human cytochrome P45Os Biochem Pharmacol 46, 787–790.

    Article  PubMed  CAS  Google Scholar 

  6. Edwards, R. J., Murray, B. P., Singleton, A. M., Murray, S., Davies, D S., and Boobis, A R. (1993) Identification of the epttope of an anti-peptide antibody which binds to CYPlA2 in many species including man Biochem Pharmacol 46, 213–220

    Article  PubMed  CAS  Google Scholar 

  7. Edwards, R. J., Smgleton, A. M, Murray, B P., Davies, D. S, and Boobts, A. R (1995) Short synthetic peptides exploited for reliable and specific targeting of antibodies to the C-termmi of cytochrome P450 enzymes. Biochem Pharmacol. 49, 39–47.

    Article  PubMed  CAS  Google Scholar 

  8. Anilkumar, T. V., Goidmg, M., Edwards, R. J., Lalani, E., Sarraf, C. E, and Alison, M, R. (1995) The resistant hepatocyte model of carcmogenesrs in the rat the apparent Independent development of oval cell proliferation and early nodules. Carcinogenesis 16, 845–853.

    Article  PubMed  CAS  Google Scholar 

  9. Goldmg, M., Sarraf, C. E., Lalam, E., Anilkumar, T V., Edwards, R J, Nagy, P, Thorgeirsson, S. S., and Ahson, M R. (1995) Oval cell differentiation mto hepatocytes in the acetylaminofluorene-treated regenerating rat liver. Hepatologv 22, 1243–1253.

    Google Scholar 

  10. Edwards, R. J, Singleton, A. M., Murray, B. P., Murray, S., Boobts, A R, and Davies, D. S. (1991) Identification of a functionally conserved surface region of rat cytochromes P450IA. Biochem. J 278, 749–757

    PubMed  CAS  Google Scholar 

  11. Edwards, R. J., Singleton, A M., Murray, B. P., Sesardic, D., Rich, K J, Davies, D. S., and Boobis, A. R(1990) An anti-peptide antibody targeted to a specrtic region of rat cytochrome P450IA2 inhibits enzyme activity. Biochem J. 266, 497–504.

    PubMed  CAS  Google Scholar 

  12. Adams, D. A., Edwards, R. J., Davies, D S., and Boobis, A R. (1997) Specific inhibition of human CYPlA2 using a targeted antibody. Biochem. Pharm 54, 189–197

    Article  PubMed  CAS  Google Scholar 

  13. Rich, K J, Sesardic, D, Foster, J R, Davies, D. S., and Boobis, A. R. (1989) Immunohlstochemical localization of cytochrome P450b/e in hepatlc and extrahepatic tissues of the rat. Biochem Pharmacol. 38, 3305–3322.

    Article  PubMed  CAS  Google Scholar 

  14. Debra, K., Boobis, A. R., Davies, D. S, and Edwards, R J (1995) Dlstributton and mduction of CYP3Al and CYP3A2 in rat liver and extra-hepattc tissues Biochem Pharmacot 50, 2047–2056.

    Article  Google Scholar 

  15. Edwards, R. J., Murray, B. P., and Boobis, A R. (1991) Anti-pepttde antibodies in studies of cytochromes P450IA. Methods Enzymol 206, 220–233

    Article  PubMed  CAS  Google Scholar 

  16. Edwards, R. J., Murray, B. P., Boobis, A. R., and Davies, D. S (1989) Identification and location of alpha-hehces in mammalian cytochromes P450. Biochemzstry 28, 3762–3770

    Article  CAS  Google Scholar 

  17. Edwards, R. J, Murray, B. P., Singleton, A. M., and Boobis, A. R (1991) Orientation of cytochromes P450 in the endoplasmic reticulum. Biochemwtry 30, 7 l–76.

    Google Scholar 

  18. Pearson, W. R. and Miller, W. (1992) Dynamic programming algorithms for biological sequence comparison Methods Enzymol. 210, 575–601.

    Article  PubMed  CAS  Google Scholar 

  19. Pennmgton, M W and Dunn, B. M. (1994) Peptide Syntheses Protocols Humana, Totowa.

    Book  Google Scholar 

  20. Atherton, E. and Sheppard, R C. (1989) Solid phase peptide synthesis—a practical approach. IRL Press, Oxford University, Oxford, UK.

    Google Scholar 

  21. Edwards, R. J., Singleton, A. M., Sesardic, D., Boobis, A R., and Davies, D. S (1988) Antibodies to a synthetic peptide that react specrfically with a common surface region on to hydrocarbon-inducible isoenzymes of cytochrome P-450 Biochem. Pharmacol 37, 3735–3741.

    Article  PubMed  CAS  Google Scholar 

  22. Edwards, R. J., Murray, B. P, Murray, S, Schulz, T., Neubert, D., Gant, T W, Thorgeirsson, S. S., Boobis, A. R., and Davies, D S (1994) Contribution of CYPl Al and CYPlA2 to the activation of heterocychc ammes in monkeys and human. Carclnogenesis 15, 829–836.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Section on Clinical Pharmacology, Imperial College School of Medicine, London, UK

    Robert J. Edwards

Authors
  1. Robert J. Edwards
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Department of Biochemistry, Queen Mary and Westfie1d College, University of London, UK

    Ian R. Phillips

  2. Department of Biochemistry and Molecular Biology, University College, London, UK

    Elizabeth A. Shephard

Rights and permissions

Reprints and permissions

Copyright information

© 1998 Humana Press Inc.

About this protocol

Cite this protocol

J. Edwards, R. (1998). Targeting Antipeptide Antibodies Towards Cytochrome P450 Enzymes. In: Phillips, I.R., Shephard, E.A. (eds) Cytochrome P450 Protocols. Methods in Molecular Biology™, vol 107. Humana Press. https://doi.org/10.1385/0-89603-519-0:239

Download citation

  • DOI: https://doi.org/10.1385/0-89603-519-0:239

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-519-5

  • Online ISBN: 978-1-59259-580-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation