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.
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References
Cregg, J. M. and Madden, K. R. (1988) Development of the methylotrophic yeast Pichia pastoris as a host for the production of foreign proteins. Dev. Ind. Microbiol. 29, 33–41.
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© 2007 Humana Press Inc., Totowa, NJ
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Ríos, S.E., Giaccone, E.M., Gerngross, T.U. (2007). Rapid Screening of Chromatography Resins for the Purification of Proteins. In: Cregg, J.M. (eds) Pichia Protocols. Methods in Molecular Biology, vol 389. Humana Press. https://doi.org/10.1007/978-1-59745-456-8_7
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DOI: https://doi.org/10.1007/978-1-59745-456-8_7
Publisher Name: Humana Press
Print ISBN: 978-1-58829-429-6
Online ISBN: 978-1-59745-456-8
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