Selection of recombinant antibodies by eukaryotic ribosome display

Methods Mol Biol. 2008:484:193-205. doi: 10.1007/978-1-59745-398-1_13.

Abstract

Ribosome display is a powerful method for selection of single-chain antibodies in vitro. It operates through the formation of libraries of antibody-ribosome-mRNA complexes that are selected on immobilized antigen, followed by recovery of the genetic information from the mRNA by RT-PCR. Both prokaryotic and eukaryotic versions are used. We describe our eukaryotic system, in which rabbit reticulocyte extracts are used for cell free transcription/translation and cDNA is recovered by in situ RT-PCR performed on the selected complexes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies / chemistry*
  • Eukaryotic Cells / chemistry*
  • Humans
  • Polymerase Chain Reaction / methods
  • Rabbits
  • Recombinant Proteins / chemistry*
  • Recombinant Proteins / genetics
  • Ribosomes / chemistry*

Substances

  • Antibodies
  • Recombinant Proteins