Expression and purification of recombinant antibody formats and antibody fusion proteins

Methods Mol Biol. 2014:1131:273-95. doi: 10.1007/978-1-62703-992-5_18.

Abstract

In the laboratory-scale production of antibody fragments or antibody fusion proteins, it is often difficult to keep track on the most suitable affinity tags for protein purification from either prokaryotic or eukaryotic host systems. Here, we describe how such recombinant proteins derived from Escherichia coli lysates as well as HEK293 cell culture supernatants are purified by IMAC and by different affinity chromatography methods based on fusions to FLAG-tag, Strep-tag, and Fc domains.

MeSH terms

  • Antibodies / genetics
  • Antibodies / metabolism*
  • Cell Line
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Humans
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism*

Substances

  • Antibodies
  • Recombinant Fusion Proteins
  • Recombinant Proteins