Screening Phage-Display Antibody Libraries Using Protein Arrays

Methods Mol Biol. 2018:1701:365-380. doi: 10.1007/978-1-4939-7447-4_20.

Abstract

Phage-display technology constitutes a powerful tool for the generation of specific antibodies against a predefined antigen. The main advantages of phage-display technology in comparison to conventional hybridoma-based techniques are: (1) rapid generation time and (2) antibody selection against an unlimited number of molecules (biological or not). However, the main bottleneck with phage-display technology is the validation strategies employed to confirm the greatest number of antibody fragments. The development of new high-throughput (HT) techniques has helped overcome this great limitation. Here, we describe a new method based on an array technology that allows the deposition of hundreds to thousands of phages by micro-contact on a unique nitrocellulose surface. This setup comes in combination with bioinformatic approaches that enables simultaneous affinity screening in a HT format of antibody-displaying phages.

Keywords: Antibodies; Array; High-throughput screening; Phage display; scFv.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cloning, Molecular / methods*
  • Humans
  • Peptide Library*
  • Protein Array Analysis / methods*
  • Single-Chain Antibodies / chemistry*
  • Single-Chain Antibodies / genetics*

Substances

  • Peptide Library
  • Single-Chain Antibodies