Rapid screening of chromatography resins for the purification of proteins

Methods Mol Biol. 2007:389:99-106. doi: 10.1007/978-1-59745-456-8_7.

Abstract

With an ever increasing number of proteins being expressed in the Pichia system, there is a growing need to rapidly develop scalable and robust purification schemes. This chapter describes a high-throughput method to screen for the optimal chromatography conditions and resin to capture and release a protein secreted by Pichia pastoris. The method involves a chromatography matrix involving four resins (Q-Sepharose, DEAE-Sepharose, SP-Sepharose, and CMSepharose), 4 pHs from 5.0 to 8.0, and 3 NaCl concentrations. The method was tested on three proteins and found to be reproducible and easily scalable.

MeSH terms

  • Chromatography, Ion Exchange
  • Enzyme-Linked Immunosorbent Assay
  • Fungal Proteins / isolation & purification*
  • Pichia / metabolism*
  • Resins, Synthetic / metabolism*

Substances

  • Fungal Proteins
  • Resins, Synthetic