Evaluation of strategies to control Fab light chain dimer during mammalian expression and purification: A universal one-step process for purification of correctly assembled Fab

Biotechnol Bioeng. 2015 Jul;112(7):1472-7. doi: 10.1002/bit.25550. Epub 2015 Mar 12.

Abstract

Fabs are an important class of antibody fragment as both research reagents and therapeutic agents. There are a plethora of methods described for their recombinant expression and purification. However, these do not address the issue of excessive light chain production that forms light chain dimers nor do they describe a universal purification strategy. Light chain dimer impurities and the absence of a universal Fab purification strategy present persistent challenges for biotechnology applications using Fabs, particularly around the need for bespoke purification strategies. This study describes methods to address light chain dimer formation during Fab expression and identifies a novel CH 1 affinity resin as a simple and efficient one-step purification for correctly assembled Fab.

Keywords: antibody fragment; expression; fab; light chain dimer; purification.

MeSH terms

  • Dimerization
  • Gene Expression*
  • Immunoglobulin Fab Fragments / biosynthesis*
  • Immunoglobulin Fab Fragments / genetics
  • Immunoglobulin Fab Fragments / isolation & purification*
  • Immunoglobulin Light Chains / biosynthesis*
  • Immunoglobulin Light Chains / genetics
  • Immunoglobulin Light Chains / isolation & purification*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics

Substances

  • Immunoglobulin Fab Fragments
  • Immunoglobulin Light Chains
  • Recombinant Proteins