Rapid screening of chromatography resins for the purification of proteins.
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.
Ríos
Sandra E
SE
GlycoFi Inc., Lebanon, NH, USA.
Giaccone
Erin M
EM
Gerngross
Tillman U
TU
D002852:Chromatography, Ion Exchange; D004797:Enzyme-Linked Immunosorbent Assay; D005656:Fungal Proteins / Q000302:isolation & purification*; D010843:Pichia / Q000378:metabolism*; D012117:Resins, Synthetic / Q000378:metabolism*
D005656:Fungal Proteins; D012117:Resins, Synthetic