| Based on short chromatographic column theory, a chromatographic pie with diameter being 500mm in productive-scale was designed and manufactured. To prove the chromatographic pie in semi-preparative scale to have good separation efficiency, a same geometric volume of an usual chromatographic column was employed to compare to the chromatographic pie from each other. In addition, the new productive technology of the simultaneous renaturation and purification of the recombinant human interferon- Y (rhIFN- Y) presented in the thesis was compared to the traditional one which is usually composed of two separate steps of renaturation and purification. On condition that according to the required purity and specific activity, the recoveries of mass and bioactivity and the purity of the rhIFN- Y obtained from the new technology and traditional one were also compared. Furthermore, the time of the productive cycle and cost were evaluated. From these foregoing results it indicates that the new technology was superior to the traditional one. The thesis includes three parts as following: 1. Principle of short chromatographic columnThe retention behavior of biopolymers was mainly controlled by the displacing agent concentration in the mobile phase employed. Thus, the separation efficiency of biopolymers was almost independent of column length . Based on the resolution for solute separation increases with the decreases in the diameter of the packed particles, the chromatographic pie was packed with particles having average diameter 5 ^m for preparative purpose. The chromatographic pie thus has very low back pressure, even though under the condition of high flow rate of mobile phase, resulting to shorten the productive cycle and lower the cost. An equation of the minimum length of column was theoretically derived and the minimum column length for separating some biopolymer was also calculated under a certain chromatographic condition. Thecalculated results were tested by experiments and the experimental results fit those equations well. It bases the strongly theoretical basis for designing and manufacturing the preparative chromatographic pie.2. Design and manufacture of 50 X SOOmml.D. chromatographic pieThe chromatographic pie with 50 X SOOmml.D companying with its support truck for productive purpose was designed. The designed principles of the productive scale chromatographic pie, shell materials, sealing method, the distribution of mobile phase and its overall structure were also elucidated. The packings with diameter Sum were packed into the chromatographic pie with radial high-pressure. Because the chromatographic pie was packed and sometimes operated under a high-pressure condition, how to choose the materials and thickness of wall of the shell of the chromatographic pie must be thought over. By means of theoretical evaluating, the structure of chromatographic pie and thickness of stainless steel were optimized.3. New technology of simultaneous renaturation and purification of recombinant human interferon- YTraditional technology of renaturation and purification of the rhIFN- Y is that the extract of rhIFN- Y from E. Coll 7.0 mol/L guanidine hydrochloride solution is firstly renaturation by diluting, then it is purified with several separation steps. By means of following the report from reference, three kinds of chromatographic steps, ion exchange chromatography, immobilized metal ion affinity chromatography and size exclusion chromatography must be employed to obtain the purity of the rhIFN- Y over 95%, as long as the inclusion body of the rhIFN- Y is high enough. However, it took 20 hours to accomplish the whole renaturation and purification of the rhIFN- Y to obtain purity of rhIFN- Y being only 90% in this study, because the'content of the rhIFN- Y in the extract was only -30 %. By using the new technology, the same extract of rhIFN- Y in guanidine hydrochloride solution was directly injected into the chromatographic pie which packed with hydrophobic hydrophobic interaction chromatographic packings...
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