Applications of hydroxyapatite-based immobilization metal affinity adsorbents for the purification of recombinant proteins
碩士 === 國立中興大學 === 化學工程學系 === 91 === In this study, the application of hydroxyapatite as IMAC absorbent for the purification of soluble recombinant poly(His)-tagged D-hydantoinase and 2-epimerase was investigated. Fe3+ as the coordinating metal ion showed the best adsorption selectivity. The...
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| Format: | Others |
| Language: | zh-TW |
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2003
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| Online Access: | http://ndltd.ncl.edu.tw/handle/60791751453285498230 |
| Summary: | 碩士 === 國立中興大學 === 化學工程學系 === 91 === In this study, the application of hydroxyapatite as IMAC absorbent for the purification of soluble recombinant poly(His)-tagged D-hydantoinase and 2-epimerase was investigated. Fe3+ as the coordinating metal ion showed the best adsorption selectivity. The addition of 0.15~0.25M NaCl and 0.05M NaF can increase D-hydantoinase recovery and activity. Changes in pH and concentration of sodium phosphate buffer can wash nonspecifically adsorbed proteins and elute D-hydantionase from adsorbent effectively. The recovery yield and purity of D-hydantoinase were 40% and above 95% respectively, under optimal purification conditions. The capacity of hydroxyapatite-based IMAC absorbent was 9.92 mg/g HAP for total proteins, and 4.9 mg/g HAP for D-hydantoinase, The absorption of denatured 2-epimerase demonstrate that hydroxyapatite-based IMAC absorbent can be applied to purification of recombinant poly(His)-tagged protein under denaturing conditions.
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