Cell-free ribosome display and selection of antibodies on arrayed antigens

Proteomics. 2016 Apr;16(8):1291-6. doi: 10.1002/pmic.201500412. Epub 2016 Apr 4.

Abstract

In vitro display technology is a powerful tool for discovery and optimisation of novel antibodies. With increasing demands on various binding molecules in proteomics studies, techniques for a large-scale generation of antibodies or antibody fragments are needed. Here, we describe a novel method for parallel generation of different antibody fragments (scFv) by integrating cell-free ribosome display with array technology. We have demonstrated the procedure by successfully isolating scFv antibodies specific to 16 different cancer biomarkers via a single process. Our results provide proof of principle for multiple production of various scFv antibodies simultaneously.

Keywords: Cell free ribosome display; Dot blot screening; Multiple antibody generation in vitro; Protein arrays.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antibody Specificity / immunology
  • Antigens / genetics
  • Antigens / immunology
  • Antigens / metabolism*
  • Biomarkers, Tumor / genetics
  • Biomarkers, Tumor / immunology
  • Biomarkers, Tumor / metabolism
  • Blotting, Western
  • Cell-Free System
  • Enzyme-Linked Immunosorbent Assay
  • Escherichia coli / genetics
  • Humans
  • Neoplasms / genetics
  • Neoplasms / immunology
  • Neoplasms / metabolism*
  • Peptide Library
  • Protein Array Analysis / methods*
  • Protein Binding / immunology
  • Protein Biosynthesis
  • Recombinant Proteins / immunology
  • Recombinant Proteins / metabolism
  • Reproducibility of Results
  • Ribosomes / genetics
  • Ribosomes / metabolism*
  • Sequence Homology, Amino Acid
  • Single-Chain Antibodies / genetics
  • Single-Chain Antibodies / immunology
  • Single-Chain Antibodies / metabolism*

Substances

  • Antigens
  • Biomarkers, Tumor
  • Peptide Library
  • Recombinant Proteins
  • Single-Chain Antibodies