Rapid Screening of Chromatography Resins for the Purification of Proteins
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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 CM-Sepharose),
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.
Affiliation(s): (2) GlycoFi Inc., Lebanon, NH
(3) Thayer School of Engineering and the Department of Biological Sciences, Dartmouth College, Hanover, NH
(3) Thayer School of Engineering and the Department of Biological Sciences, Dartmouth College, Hanover, NH
Book Title: Pichia Protocols
Series: Methods in Molecular Biology | Volume: 389 | Pub. Date: Aug-08-2007 | Page Range: 99-106 | DOI: 10.1007/978-1-59745-456-8_7
Subject: Biotechnology
Key Words: Pichia pastoris - protein purification - high-throughput screening - yeast - ion exchange chromotography resins
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