科研之友
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摘要
This research aimed to exercise a large scale (bioreactor) process based on a laboratory experiment and to get control of process parameters on a large scale (bioreactor) to prepare an protein interferon 汐-2b. The study was initiated by a laboratory-scale protein interferon 汐-2b overproduction, as a preparation for performing a bioreactor process scale-up. Bioreactor scale-up was performed on a scale of 10 L, based on oxygen mass transfer coefficients and kL汐 values between laboratory scale and bioreactor scale. To determine the kL汐 , an optimization process was carried out considering a variety of important variables affecting the process in a bioreactor scale. Subsequently, the obtained parameters in the form of aeration velocity and process control were selected. Further, an optimization was also by varying the fermentation time at 40, 60, 72, 80 and 96 h, to get the highest yield of substrate and the protein. Our experiment showed that the kL汐 (oxygen mass transfer coefficient), incubation time and aeration velocity were the most important parameters influencing the up production process of the protein up to 10x scaling up. Protein purification was performed by using a nickel affinity column chromatography, while purified protein was analyzed by SDS-PAGE. The quality of interferon resulted from bioreactor scale was similar to that of laboratory scale.
