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Rapid Screening of Chromatography Resins for the Purification of Proteins
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 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
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
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