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Biotechnol J ; 7(10): 1221-32, 2012 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-22887918

RESUMEN

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.


Asunto(s)
Cromatografía por Intercambio Iónico/métodos , Ensayos Analíticos de Alto Rendimiento/métodos , Proteínas de la Leche/aislamiento & purificación , Modelos Químicos , Adsorción , Resinas de Intercambio Aniónico/química , Precipitación Química , Cromatografía de Afinidad/instrumentación , Cromatografía de Afinidad/métodos , Cromatografía por Intercambio Iónico/instrumentación , Interacciones Hidrofóbicas e Hidrofílicas , Modelos Lineales , Reproducibilidad de los Resultados , Cloruro de Sodio/química , Proteína de Suero de Leche
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