Development of Loofa Sponge-Based Immobilized Metal Affinity Chromatography Adsorbents
碩士 === 國立中興大學 === 化學工程學系所 === 106 === In this study, loofa-sponge was as a support, the use of hydroxyethyl cellulose cross-linked to the substrate surface to increase its affinity ligand, the use of chemical synthesis of epichlorohydrin and chelator iminodiacetic acid and transition metal ions, fin...
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| Format: | Others |
| Language: | zh-TW |
| Published: |
2018
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| Online Access: | http://ndltd.ncl.edu.tw/handle/p8q9mq |
| Summary: | 碩士 === 國立中興大學 === 化學工程學系所 === 106 === In this study, loofa-sponge was as a support, the use of hydroxyethyl cellulose cross-linked to the substrate surface to increase its affinity ligand, the use of chemical synthesis of epichlorohydrin and chelator iminodiacetic acid and transition metal ions, finally after the immobilized metal affinity chromatography adsorption substrate is completed, the transition metal ions could be reacted to coordinate covalent bonds with hexahistidine-Tag on the genetically modified Picrophilus torridus trehalose synthase to purify and immobilize the trehalose synthase.
From the results of this study, we obtained the most suitable bifunctional support for the target protein with both the IMAC-adsorbed enzyme terminus and the epoxy functional group covalently bonded to the enzyme. The kinetics of adsorption and incubation time experiments established the absoption time and the immobilized time. The stability of the physical adsorption and covalent bonding was finally explored to confirm that the covalent bonding is more stable than the physical adsorption.
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