Purification of Recombinant Glycoproteins from Pichia pastoris Culture Supernatants

Methods Mol Biol. 2019:1923:343-350. doi: 10.1007/978-1-4939-9024-5_17.

Abstract

Pichia pastoris is a common host organism for the production of recombinant proteins. While unglycosylated recombinant proteins derived from this yeast can be purified efficiently by only a few conventional chromatography steps, the purification of glycosylated recombinant proteins is a very challenging process due to the intrinsic feature of the yeast of hypermannosylation. The resulting vast glycosylation pattern on the recombinant target protein masks its physicochemical properties hampering a conventional downstream process. Here, we describe a fast and efficient two-step chromatography strategy, where both steps are operated in flow-through mode, to purify recombinant glycoproteins from P. pastoris culture supernatants.

Keywords: Downstream process; Flow-through chromatography; Glycosylation; Pichia pastoris; Recombinant protein production.

MeSH terms

  • Chromatography
  • Glycoproteins / genetics
  • Glycoproteins / metabolism*
  • Glycosylation
  • Pichia / genetics
  • Pichia / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism*

Substances

  • Glycoproteins
  • Recombinant Proteins