| The objectives of this research were to optimize the yield and purity of heterologous proteins from a complex, polydisperse suspension like transgenic milk by microfiltration (MF), ultrafiltration (UF) and expanded bed adsorption chromatography (EBA) and to develop a global model to predict general MF/UF processes individually or in any combination. This was achieved by a synergistic research strategy that incorporated MF (lab-scale followed by pilot plant) to optimize yield as the first step. In the second step, to optimize purity, a two track approach---UF and EBA was adopted. A two-pronged method---optimum fluid dynamics and optimum electrostatics was employed to obtain high yields and purity. In parallel, modeling of the filtration processes was developed with the ultimate objective of developing a global predictive algorithm supported by a computer program to simulate and optimize MF/UF filtration processes. This resulted in a facile methodology which can simulate and optimize a wide variety of MF/UF processes enabling in-silico experiments for MF/UF applications for various suspensions, membrane sizes and solution conditions like pH, ionic strength and temperature. MF/UF processes can thus be optimized to minimize operating time, maximize purity of a product or to maximize the yield of a product. Thus, this research encompasses various facets of membrane technology: helical Dean vortices module, different membrane chemistries like polyether sulfone, regenerated cellulose and ceramic, various membrane schemes, advanced operations involving pH, ionic strength and back-pulsing and new theoretical developments to predict and optimize MF/UF processes a priori. Additionally a competing novel chromatographic technology (EBA) was investigated as an alternative to UF for protein purification. |
