High-throughgput phage-display screening in array format

Enzyme Microb Technol. 2015 Nov:79-80:34-41. doi: 10.1016/j.enzmictec.2015.06.016. Epub 2015 Jul 14.

Abstract

Emerging technologies for the design and generation of human antibodies require improved approaches enabling their screening, characterization and validation. Currently, strategies based on ELISA or western blot are used to that aim. However, the ever increasing number of novel antibodies generated would benefit from the development of new high-throughput (HT) platforms facilitating rapid antibody identification and characterization. Herein, we describe a protein chip bearing recombinant phage particles and based on a large phage antibody library. In this paper we have set forth a novel implementation which provides a powerful and simple methodology enabling the identification of single-chain variable fragments (scFv). As a proof-of-principle of this method, we tested it with recombinant antigen (human recombinant interleukin 8). Additionally, we developed a novel bioinformatics tool that serves to compare this novel strategy with traditional methods. The method described here, together with associated informatics tools, is robust, relatively fast and represents a step-forward in protocols including phage library screenings.

Keywords: Antibody; Chip; High-throughput screening; Phage library; Phage-display array.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Cell Surface Display Techniques / methods*
  • Cloning, Molecular
  • Computational Biology
  • DNA / genetics
  • High-Throughput Screening Assays / methods*
  • Humans
  • Immobilized Proteins / genetics
  • Immobilized Proteins / immunology
  • Interleukin-8 / genetics
  • Interleukin-8 / immunology
  • Molecular Sequence Data
  • Peptide Library*
  • Protein Array Analysis / methods
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Single-Chain Antibodies / chemistry
  • Single-Chain Antibodies / genetics

Substances

  • CXCL8 protein, human
  • Immobilized Proteins
  • Interleukin-8
  • Peptide Library
  • Recombinant Proteins
  • Single-Chain Antibodies
  • DNA