Determination of adsorption isotherm parameters for minor whey proteins by gradient elution preparative liquid chromatography.
Ion-Exchange Chromatography (IEC) techniques have been extensively investigated in protein purification processes, due to the more selective and milder separation steps. To date, existing studies of minor whey proteins fractionation in IEC have primarily been conducted as batch uptake studies, which require more experimental search space, time and materials. In this work, the selected resin's (SP Sepharose FF) equilibrium and dynamic binding capacity were first investigated. Next, adsorption of the pure binary mixture of lactoperoxidase and lactoferrin was studied to calibrate steric mass action (SMA) model using a simplified approach with data from single column experiments. The calibrated model was then verified by performing factorial-design based experiments for various process operating conditions assessing process performance on a larger bed height column. The model predicted results demonstrated a realistic agreement with the experiments providing reproducible column elution profile and reduced experimental work. Finally, whey protein isolate was used to evaluate model parameters in real conditions. Results obtained herein are suitable for future large scale applications.
Department of Chemical and Biochemical Engineering, University of Western Ontario, London, Ontario, Canada.
Naeimeh
N
Faraji
Faculty of Engineering and Applied Science, Memorial University of Newfoundland, St John's, Newfoundland and Labrador, Canada.
Yan
Y
Zhang
Department of Chemical and Biochemical Engineering, University of Western Ontario, London, Ontario, Canada. Electronic address: aray@eng.uwo.ca.
Ajay K
AK
Ray
D000327:Adsorption; D002852:Chromatography, Ion Exchange / Q000379:methods; D007781:Lactoferrin / Q000032:analysis; D012685:Sepharose; D000067816:Whey Proteins / Q000032:analysis*
D000067816:Whey Proteins; D012685:Sepharose; D007781:Lactoferrin
Adsorption isotherm; Downstream process development; Inverse method; Mathematical modeling; Milk protein separation; Steric mass action; Whey proteins