Adaptation of hybridomas to protein-free media results in a simplified two-step immunoglobulin M purification process

J Chromatogr A. 2009 Mar 27;1216(13):2683-8. doi: 10.1016/j.chroma.2008.10.067. Epub 2008 Oct 21.

Abstract

An IgM antibody was purified from hybridoma supernatant containing serum using a three-step purification process comprising of tangential flow filtration, anion-exchange chromatography and size-exclusion chromatography. Recovery and purity were significantly improved upon adaptation of the hybridoma to serum-free media. The process could even be simplified by omitting the initial tangential flow filtration step. Even with a two-step purification process a purity of >98% and a recovery of >60% was obtained.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatography, Gel
  • Chromatography, Ion Exchange
  • Culture Media
  • Culture Media, Serum-Free
  • Hybridomas / immunology*
  • Immunoglobulin M / isolation & purification*
  • Mice

Substances

  • Culture Media
  • Culture Media, Serum-Free
  • Immunoglobulin M