Monitoring Phage Biopanning by Next-Generation Sequencing

Methods Mol Biol. 2018;1701:463-473. doi: 10.1007/978-1-4939-7447-4_26.

Abstract

Phage display has enabled the rapid isolation of antigen-specific antibodies from combinatorial libraries of the variable heavy chain gene (VH) and variable light chain gene (VL). The method is based on genetic engineering of bacteriophages and repeated rounds of antigen-guided selection by phage biopanning.Next-Generation Sequencing (NGS) coupled with bioinformatics are powerful tools for analyzing the large number of DNA sequences present in an immune library.Here, we describe a method that demonstrates how NGS analysis enhances phage biopanning of complex antibody libraries as well as facilitates the antibody discovery process.

Keywords: Antibody repertoire; B-Cell receptor repertoire; HTS; High-throughput sequencing; Immunoglobulin repertoire; NGS; Next-Generation Sequencing; V gene; VDJ; Variable region; scFv.

MeSH terms

  • Animals
  • Cloning, Molecular / methods*
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Peptide Library*
  • Single-Chain Antibodies / genetics*

Substances

  • Peptide Library
  • Single-Chain Antibodies