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題名 | Metal-chelating Pluronics for Immobilization of Histidine-tagged Proteins at Interfaces:Firefly Luciferase on the Polystyrene Beads=金屬螯合 Pluronics[feb9] 聚醚醇於組胺酸-蛋白質固定化之應用 |
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作者 | Ho,Chih-hu; Limberis,Loren; Stewart,Russell J.; |
期刊 | 界面科學會誌 |
出版日期 | 19971200 |
卷期 | 20:2 1997.12[民86.12] |
頁次 | 頁95-106 |
分類號 | 467.4 |
語文 | eng |
關鍵詞 | 金屬螯合; 聚醚醇; 組胺酸; 蛋白質固定化; |
英文摘要 | A convenient and general method of protein immobilization was developed that takes advantage of the self-organizing and protein-repelling properties of PluronicsR surfactants at hydrophobic interfaces. The activity preserving PluronicsR interface was used as a foundation for the specific immobilization of histidine-tagged proteins through their affinity for immobilized metal ions. PluronicsR F108 was modified with the chelating group Nitrilotriacetic Acid (NTA) to form F 1 08-NTA as the surface modification reagent. Histidine-tagged firefly luciferase (FFL), the model protein, and polystyrene (PS) beads (0.453 um diameter), the model surface, were used to examine the specific metal affinity immobilization and protein repelling properties of F108-NTA. We found that the FFL specifically attached to the F108-NTA modified PS beads through chelated Ni2+ions. In addition, we found that the FFL retained approximately 95% of its bioluminescent activity while bound to the surface. |
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