Application of hydrophobic interaction displacement chromatography for an industrial protein purification.
Recently it has been established that low molecular weight displacers can be successfully employed for the purification of proteins in hydrophobic interaction chromatography (HIC) systems. This work investigates the utility of this technique for the purification of an industrial protein mixture. The study involved the separation of a mixture of three protein forms, that differed in the C-terminus, from their aggregate impurities while maintaining the same relative ratio of the three protein forms as in the feed. A batch high-throughput screening (HTS) technique was employed in concert with fluorescence spectroscopy for displacer screening in these HIC systems. This methodology was demonstrated to be an effective tool for identifying lead displacer candidates for a particular protein/stationary-phase system. In addition, these results indicate that surfactants can be employed at concentrations above their CMCs as effective displacers. Displacement of the recombinant proteins with PEG-3400 and the surfactant Big Chap was shown to increase the productivity as compared to the existing step-gradient elution process.
Sunasara
Khurram M
KM
Department of Chemical Engineering, Rensselaer Polytechnic Institute, Troy, New York 12180, USA.
Xia
Fang
F
Gronke
Robert S
RS
Cramer
Steven M
SM
D002845:Chromatography / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D019544:Equipment Failure Analysis; D005240:Feasibility Studies; D057927:Hydrophobic and Hydrophilic Interactions; D008970:Molecular Weight; D010865:Pilot Projects; D011092:Polyethylene Glycols; D011786:Quality Control; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification*; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013501:Surface-Active Agents
D011994:Recombinant Proteins; D013501:Surface-Active Agents; D011092:Polyethylene Glycols