Construction of phage antibody repertoires from the blood of West Nile virus-infected donors

Methods Mol Biol. 2009:562:45-60. doi: 10.1007/978-1-60327-302-2_4.

Abstract

A method for the construction of West Nile virus immune donor antibody repertoires is described. B cells are harvested from a suitable donor and the antibody variable genes are amplified using polymerase chain reaction (PCR). The PCR fragments are cloned in a phage display vector to construct a repertoire that can be used in panning procedures to identify many unique monoclonal antibodies.

MeSH terms

  • Antibodies, Viral / immunology*
  • Antigens, Viral / immunology
  • Bacteriophages / immunology*
  • Blood Donors*
  • Humans
  • Immunoglobulin Heavy Chains / genetics
  • Immunoglobulin Heavy Chains / immunology
  • Immunoglobulin Light Chains / genetics
  • Immunoglobulin Light Chains / immunology
  • Immunoglobulin Variable Region / genetics
  • Immunoglobulin Variable Region / immunology
  • Peptide Library*
  • Protein Engineering
  • RNA, Viral / genetics
  • RNA, Viral / metabolism
  • West Nile Fever / blood*
  • West Nile virus / immunology*

Substances

  • Antibodies, Viral
  • Antigens, Viral
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Light Chains
  • Immunoglobulin Variable Region
  • Peptide Library
  • RNA, Viral