Model-based high-throughput process development for chromatographic whey proteins separation.
In this study, an integrated approach involving the combined use of high-throughput screening (HTS) and column modeling during process development was applied to an industrial case involving the evaluation of four anion-exchange chromatography (AEX) resins and four hydrophobic interaction chromatography (HIC) resins for the separation of whey proteins having close pIs. From the HTS data, one resin of each type was selected (Capto Q and Octyl Sepharose 4 FF). Next, batch uptake experiments were performed to determine the adsorption isotherms of the major whey proteins on the selected resins, followed by isotherm parameters regression. Using the obtained isotherm parameters, the candidate chromatographic operations were modeled and experimentally validated. Finally, these were model-optimized and evaluated based on their optimized performances. In this example, Capto Q performed much better than Octyl Sepharose 4 FF in terms of column capacity, loading, throughput and productivity; hence, Capto Q was selected as the resin of choice for whey proteins separation. This operation was further scaled up from the lab scale (1 mL) to a preparative scale (35 L), with reproducible column elution profile. By this approach, a much wider space of operating variables could be investigated, thereby increasing the chances of finding the ideal operating conditions while keeping experimentation to the minimum.
Department of Biotechnology, Delft University of Technology, Delft, The Netherlands.
Beckley K
BK
Nfor
Jovana
J
Ripić
Albert
A
van der Padt
Marc
M
Jacobs
Marcel
M
Ottens
D000327:Adsorption; D000837:Anion Exchange Resins / Q000737:chemistry; D011232:Chemical Precipitation; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000295:instrumentation / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D057927:Hydrophobic and Hydrophilic Interactions; D016014:Linear Models; D008894:Milk Proteins / Q000302:isolation & purification*; D008956:Models, Chemical; D015203:Reproducibility of Results; D012965:Sodium Chloride / Q000737:chemistry; D000067816:Whey Proteins
D000837:Anion Exchange Resins; D008894:Milk Proteins; D000067816:Whey Proteins; D012965:Sodium Chloride