Modeling and analysis of the affinity filtration process, including broth feeding, washing, and elution steps.
Affinity filtration is a developing protein purification technique that combines the high selectivity of affinity chromatography and the high processing speed of membrane filtration. In this work a lumped kinetic model was developed to describe the whole affinity filtration process, including broth feeding, contaminant washing, and elution steps. Affinity filtration experiments were conducted to evaluate the model using bovine serum albumin as a model protein and a highly substituted Blue Sepharose as an affinity adsorbent. The model with nonadjustable parameters agreed fairly to the experimental results. Thus, the performance of the affinity filtration in processing a crude broth containing contaminant proteins was analyzed by computer simulations using the lumped model. The simulation results show that there is an optimal protein loading for obtaining the maximum recovery yield of the desired protein with a constant purity at each operating condition. The concentration of a crude broth is beneficial in increasing the recovery yield of the desired protein. Using a constant amount of the affinity adsorbent, the recovery yield can be enhanced by decreasing the solution volume in the stirred tank due to the increase of the adsorbent weight fraction. It was found that the lumped kinetic model was simple and useful in analyzing the whole affinity filtration process.
Department of Biochemical Engineering, Tianjin University, Tianjin 300072, People's Republic of China.
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D000327:Adsorption; D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods*; D003198:Computer Simulation; D005374:Filtration / Q000295:instrumentation / Q000379:methods; D008956:Models, Chemical; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification*; D012685:Sepharose / Q000031:analogs & derivatives / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000378:metabolism / Q000493:pharmacokinetics
D011506:Proteins; D012710:Serum Albumin, Bovine; C029483:Blue-Sepharose; D012685:Sepharose