Antibody selection using clonal cocultivation of Escherichia coli and eukaryotic cells in miniecosystems

Proc Natl Acad Sci U S A. 2018 Jul 3;115(27):E6145-E6151. doi: 10.1073/pnas.1806718115. Epub 2018 Jun 18.

Abstract

We describe a method for the rapid selection of functional antibodies. The method depends on the cocultivation of Escherichia coli that produce phage with target eukaryotic cells in very small volumes. The antibodies on phage induce selectable phenotypes in the target cells, and the nature of the antibody is determined by gene sequencing of the phage genome. To select functional antibodies from the diverse antibody repertoire, we devised a selection platform that contains millions of picoliter-sized droplet ecosystems. In each miniecosystem, the bacteria produce phage displaying unique members of the antibody repertoire. These phage interact only with eukaryotic cells in the same miniecosystem, making phage available directly for activity-based antibody selection in biological systems.

Keywords: antibody selection; miniecosystem; phage display.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacteriophage M13* / genetics
  • Bacteriophage M13* / immunology
  • CHO Cells
  • Coculture Techniques
  • Cricetulus
  • Escherichia coli* / genetics
  • Escherichia coli* / immunology
  • Humans
  • Membrane Glycoproteins / antagonists & inhibitors*
  • Membrane Glycoproteins / immunology
  • Receptor, trkB / antagonists & inhibitors*
  • Receptor, trkB / immunology
  • Single-Chain Antibodies* / genetics
  • Single-Chain Antibodies* / immunology

Substances

  • Membrane Glycoproteins
  • Single-Chain Antibodies
  • Receptor, trkB
  • tropomyosin-related kinase-B, human