Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/3279
標題: 固定化金屬與heparin親和薄膜應用於板框型薄膜分離器之設計與研究
作者: 蔡岳樺
Tsai, Yueh-Hua
關鍵字: Immobilized Metal ion Affinity Membrane;固定化金屬親和薄膜;Heparin Affinity Membrane;Plate-and-frame Module;肝素親和薄膜;板框型薄膜分離器
出版社: 化學工程學系
摘要: 
The separation performance of immobilized metal and heparin affinity membranes with the use of plate-and-frame modules was investigated in this study. The proteins employed were lysozyme, BSA and recombinant HGF (hepatocyte growth factor). In the batch adsorption of lysozyme onto immobilized metal and heparin affinity membranes, the adsorption capacity was the largest when 1,8-diaminooctane was used as spacer arm, and the capacity increased with increasing pH. In addition, nonspecific binding was significant for immobilized metal affinity membrane, whereas it was negligible for heparin affinity membrane. The effect of nonspecific binding could be decreased by increasing salt concentration in the solution. In the flow process using plate-and-frame module, the design of two modules in tandem achieved the best lysozyme adsorption performance. In the experiments of two-protein separation, desorbed lysozyme amount was greater than BSA at pH 8 or 7, but the desorbed BSA amount was significantly increased at pH 5. Finally, when purifying HGF from insect cell supernatant in batch system, HGF was found to be adsorbed on immobilized metal affinity membrane, but failed to adsorb on heparin affinity membrane. As to the purification of HGF using two membrane modules in tandem, the analysis results indicate that HGF was not adsorbed onto any membrane.

本研究以固定化金屬與heparin親和薄膜配合板框型分離器之使用,探討lysozyme之吸附分離效果,並實際應用於自細胞培養上清液中純化重組肝細胞生長因子。在批式吸附部分,以1,8-diaminooctane為延伸臂之固定化金屬與heparin親和薄膜皆獲致最佳之lysozyme吸附效果,且lysozyme之吸附量隨pH值提高而增加。本研究所得之固定化金屬親和薄膜有極大比例之非特定性吸附現象,但heparin親和薄膜之非特定性吸附小,而非特定性吸附可藉加入適當之鹽類濃度來改善。在使用於板框型分離器之流動吸附部分,只使用單一性親和薄膜時以層疊二片heparin親和薄膜方式效果較佳,若使用多片且雙種親和薄膜時,則以串聯二組板框時可得較佳之lysozyme脫附量。此外,在加入BSA的雙蛋白質溶液之分離方面,於pH 8或7時,lysozyme有較大之吸附比例,但在pH 5時BSA之脫附量明顯提高。最後,自細胞培養上清液中純化分離肝細胞生長因子部分,批式吸附之結果顯示:肝細胞生長因子可與本研究之固定化金屬親和薄膜吸附鍵結,但對於heparin親和薄膜則無吸附現象;而以板框型分離器純化肝細胞生長因子時,脫附液之分析結果顯示無肝細胞生長因子吸附至任一種親和薄膜。
URI: http://hdl.handle.net/11455/3279
Appears in Collections:化學工程學系所

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