Selection of antibodies against a single rare cell present in a heterogeneous population using phage display

Nat Protoc. 2011 Apr;6(4):509-22. doi: 10.1038/nprot.2011.311. Epub 2011 Mar 24.

Abstract

Here we describe a new method applying phage-displayed antibody libraries to the selection of antibodies against a single identified cell on a glass slide. This is the only described method that has successfully achieved selection of antibodies against a single rare cell in a heterogeneous population of cells. The phage library is incubated with the slide containing the identified rare cell of interest; incubation is followed by UV irradiation while protecting the target cell with a minute disc. The UV light inactivates all phages outside the shielded area by cross-linking the DNA constituting their genomes. The expected yield is between one and ten phage particles from a single cell selection. The encoded antibodies are subsequently produced monoclonally and tested for specificity. This method can be applied within a week to carry out ten or more individual cell selections. Including subsequent testing of antibody specificity, a specific antibody can be identified within 2 months.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies / analysis*
  • Antibody Specificity
  • Immunologic Techniques
  • In Situ Hybridization, Fluorescence
  • Micromanipulation
  • Peptide Library*
  • Ultraviolet Rays

Substances

  • Antibodies
  • Peptide Library