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Purification of Recombinant Glycoproteins from Pichia pastoris Culture Supernatants

  • David Johannes Wurm
  • Oliver SpadiutEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1923)

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.

Key words

Pichia pastoris Recombinant protein production Glycosylation Downstream process Flow-through chromatography 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute of Chemical, Environmental and Bioscience Engineering, Research Division Biochemical Engineering, Vienna University of Technology (TU Wien)ViennaAustria

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