Selection of human antibody fragments by phage display

Nat Protoc. 2007;2(11):3001-8. doi: 10.1038/nprot.2007.448.

Abstract

Here, we describe a protocol for the selection of human antibody fragments using repertoires displayed on filamentous bacteriophage. Antigen-specific clones are enriched by binding to immobilized antigen, followed by elution and repropagation of phage. After multiple rounds of binding selection, specific clones are identified by ELISA. This article provides an overview of phage display and antibody technology, as well as detailed protocols for the immobilization of antigen, the selection of repertoires on purified or complex antigens and the identification of binders.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / genetics*
  • Antibodies, Monoclonal / metabolism
  • Antigen-Antibody Reactions
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Immunoglobulin Fragments / analysis
  • Immunoglobulin Fragments / genetics*
  • Immunoglobulin Fragments / metabolism
  • Peptide Library*
  • Recombinant Fusion Proteins / metabolism

Substances

  • Antibodies, Monoclonal
  • Immunoglobulin Fragments
  • Peptide Library
  • Recombinant Fusion Proteins