以螯合樹脂Amberlite IRC-748分離牛血紅素與牛血清蛋白混合物

Separation of Bovine Serum Albumin ( BSA ) and Bovine Hemoglobin ( BHb ) Mixtures with a Chelating Resin Amberlite IRC-748

翁小喬(Hsiao-Chiao Weng)

指導教授 ： 莊瑞鑫

元智大學/工程學院/化學工程與材料科學學系/碩士(2008年)

https://doi.org/10.6838/YZU.2008.00032

摘要

本研究應用螯合樹脂Amberlite IRC-748分離雙成份蛋白質混合溶液，實驗所使用的蛋白質為牛血清蛋白( BSA )及牛血紅素( BHb )，探討吸附pH值、溫度、添加鹽類對吸附之影響，以及吸附熱力學、雙成份蛋白質吸附分離與動力學分析，最後為脫附條件探討。
在單成份蛋白質吸附實驗，BHb及BSA在其等電點以下，吸附量很高。當溫度提高、添加鹽類，皆可使兩蛋白質吸附量增加，其中以BHb較明顯。另外，以Semi-reciprocal plot線性回歸分析，確定樹脂吸附BHb與BSA符合Langmuir isothermal model，且兩蛋白質在吸附過程的熱力學變化，均為吸熱反應。在雙成份蛋白質吸附分離實驗，吸附條件為40oC、pH 4.0、0.2 M NaCl，雙成份蛋白質可達完全分離；而40oC、pH 5.0、0.2 M NaCl時，分離效果佳。以三種競爭型吸附模式進行雙成份吸附的擬合探討，發現兩蛋白質有競爭吸附現象。在吸附動力學方面，以動力速率式擬合發現BHb及BSA的吸附反應符合二階速率式。再利用外部膜擴散及顆粒內部擴散模式進行吸附機制探討，吸附條件為pH 4.0、5.0時，BSA為外部膜擴散主導；而BHb應是先快速通過液膜層吸附至樹脂表面後，再開始產生內部孔洞擴散進而吸附，吸附機制為顆粒內部擴散主導。而脫附實驗方面，利用改變pH值的脫附策略，發現在低溫下吸附後脫附比在高溫下吸附再脫附容易。

關鍵字

蛋白質吸附 ； 牛血紅素 ； 牛血清蛋白 ； 樹脂 ； 熱力學

並列摘要

In this study, the application of chelating resin Amberlite IRC-748 as ion-exchange absorbent for separation of bovine serum albumin ( BSA ) and bovine hemoglobin ( BHb ) mixture. The effects of pH, temperature and salt concentration on single protein adsorption and thermodynamics were investigated. Furthermore, separation factor and the kinetics properties of binary protein system were also investigated with adsorption process. Finally, it was discussed with desorption conditions.
For both proteins, amount of adsorption was higher at low pH values. The results show that the amount of protein adsorbed increases as the temperature and salt concentration increase. In addition, the adsorption isotherms well fitted by the Langmuir isothermal model from the semi-reciprocal plot analysis. Utilizing Van’t Hoff relationships to analyze the experimental data for these interactions, the change in the apparent enthalpy of association for the adsorption of BSA and BHb onto IRC-748 was positive. In binary protein adsorption experiment, it could be separated completely at 40oC, pH 4.0 with 0.2 M NaCl. However, the condition at 40oC, pH 5.0 with 0.2 M NaCl showed the best adsorption selectivity. It has shown that adsorption behavior with competitive phenomenon when using the three competitive adsorption models in binary component system. In order to investigate the mechanism of adsorption and potential rate controlling step, first- and second-order kinetic equations, intraparticle diffusion model and external film mass transfer model have been used for testing experimental data. Adsorption of both proteins is best fit by second-order kinetic equation. In these adsorption conditions, the adsorption mechanism of BSA is external film diffusion controlling and BHb can be best described by intraparticle diffusion model. Aspect to desorption experiment, by using modified pH value will find that the desorption percentage of protein which adsorb at low temperature will better than high temperature.

並列關鍵字

Protein adsorption ； Hemoglobin ； Albumin ； Resin ； Thermodynamics

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