Advertisement

Probing the Antigenicity of HCV Envelope Glycoproteins by Phage Display Antibody Technology

  • Erick Giang
  • Fernando Aleman
  • Mansun LawEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1911)

Abstract

The envelope glycoproteins E1 and E2 of hepatitis C virus form a heterodimeric complex on the viral surface. They are the targets of neutralizing antibodies and are being investigated as potential vaccine antigens. Because of the high level of cysteine residues and N-glycosylation sites in the polypeptide sequences, it is technically challenging to produce pure, folded recombinant E1, E2, and E1E2 complex for downstream analysis. In this chapter, the methods we used to isolate a panel of human antibodies specific to diverse antigenic regions on the glycoproteins are discussed. The antibodies have been found to be valuable reagents for the study of HCV envelope glycoproteins, including the determination of the first E2 core domain structure.

Key words

Phage display Antibody Epitope Antigenic site Antigenic region Glycoproteins 

Notes

Acknowledgments

This work was supported by NIH grants AI079031, AI106005, AI123365, and AI123861.

References

  1. 1.
    Aghebati-Maleki L et al (2016) Phage display as a promising approach for vaccine development. J Biomed Sci 23:66CrossRefGoogle Scholar
  2. 2.
    Schirrmann T et al (2011) Phage display for the generation of antibodies for proteome research, diagnostics and therapy. Molecules 16:412–426CrossRefGoogle Scholar
  3. 3.
    Georgiou G et al (2014) The promise and challenge of high-throughput sequencing of the antibody repertoire. Nat Biotechnol 32:158–168CrossRefGoogle Scholar
  4. 4.
    Omidfar K, Daneshpour M (2015) Advances in phage display technology for drug discovery. Expert Opin Drug Discov 10:651–669CrossRefGoogle Scholar
  5. 5.
    Strohl WR (2014) Antibody discovery: sourcing of monoclonal antibody variable domains. Curr Drug Discov Technol 11:3–19CrossRefGoogle Scholar
  6. 6.
    Zhao AZ et al (2016) Phage antibody display libraries: a powerful antibody discovery platform for immunotherapy. Crit Rev Biotechnol 36:276–289CrossRefGoogle Scholar
  7. 7.
    Giang E et al (2012) Human broadly neutralizing antibodies to the envelope glycoprotein complex of hepatitis C virus. Proc Natl Acad Sci U S A 109:6205–6210CrossRefGoogle Scholar
  8. 8.
    Barbas Iii CF, Wagner J (1995) Synthetic human antibodies: selecting and evolving functional proteins. Methods 8:94–103CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Immunology and MicrobiologyThe Scripps Research InstituteLa JollaUSA
  2. 2.Navega Therapeutics IncLa JollaUSA

Personalised recommendations