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

Methods Mol Biol. 2019:1911:381-393. doi: 10.1007/978-1-4939-8976-8_26.

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.

Keywords: Antibody; Antigenic region; Antigenic site; Epitope; Glycoproteins; Phage display.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Bacteriophages / genetics
  • Bacteriophages / metabolism
  • Cell Surface Display Techniques / instrumentation
  • Cell Surface Display Techniques / methods*
  • Epitopes / immunology
  • Hepacivirus / immunology*
  • Hepacivirus / metabolism
  • Hepatitis C / blood
  • Hepatitis C / immunology
  • Hepatitis C / prevention & control
  • Hepatitis C Antibodies / immunology
  • Hepatitis C Antibodies / isolation & purification*
  • Humans
  • Immunogenicity, Vaccine
  • Protein Domains
  • Viral Envelope Proteins / chemistry
  • Viral Envelope Proteins / immunology*
  • Viral Hepatitis Vaccines / immunology
  • Viral Hepatitis Vaccines / therapeutic use

Substances

  • E1 protein, Hepatitis C virus
  • Epitopes
  • Hepatitis C Antibodies
  • Viral Envelope Proteins
  • Viral Hepatitis Vaccines
  • glycoprotein E2, Hepatitis C virus