Selectivity of IMAC Columns in Trypsin Inhibitor Purification
详细信息 查看全文
- 作者：Haydee Yeomans-Reina ; Arturo Ruiz-Manriquez ; Benjamin Ramirez Wong ; and Armando Tejeda Mansir
- 刊名：Biotechnology Progress
- 出版年：2001
- 出版时间：August 2001
- 年：2001
- 卷：17
- 期：4
- 页码：729 - 733
- 全文大小：60K
- 年卷期：v.17,no.4(August 2001)
- ISSN：1520-6033
文摘
The properties of an adsorbent and the parameters in an adsorption process affectthe resolution of chromatographic purifications. This is reflected in the elution profile,which shows the relative affinity of different proteins for a specific adsorbent. In thework presented here, elution profiles for trypsin inhibitor were used to study the effectsof the concentration of trypsin inhibitor, ionic strength of the protein solution, slopeof the elution gradient, and the regeneration treatment of the chromatography columnon the selectivity of the adsorbent Cellufine Chelate-CuII(ida). Cytochrome c was usedas a reference protein. Variations in the concentrations of trypsin inhibitor and in theionic strength of the buffered solution did not have any effects on the elution profile.On the other hand, changes in the slope of the pH gradient used for elution causedshifting of the elution peaks toward lower values of the elution volume, resulting inthe best strategy to modify the elution profile of the system. Finally, using a constantslope pH gradient of elution, the variation of the selectivity of the adsorbent for trypsininhibitor when subjected to cleaning treatments with 0.5 N NaOH was studied.Appropriate cleaning practices used in industry were followed. The adsorbent showedonly a slight tendency for resolution loss in the order of 2 × 10-4 days-1. The resultspresented here show a good stability of the adsorbent when compared to otherbiospecific adsorbents commonly used.