逆沖提法於吸附型管柱層析蛋白質純化之探討

The Study of Protein Purification in Adsorption Column Chromatography by Back Flush

張慧蘭(Hui-Lan Chang)

指導教授 ： 陳銘凱

淡江大學/理學院/化學學系碩士班/碩士(2012年)

https://doi.org/10.6846/TKU.2012.00610

摘要

利用帶有YNR029Cp、ALD4p基因的質體轉形至大腸桿菌（E.coli）的表現宿主細胞BL21（DE3）表現並純化後，以Q SepharoseTM Fast Flow、Hydroxylapatite（使用純化之YNR029Cp）、Phenyl Sepharose 6 Fast Flow、Blue Sepharose 6 Fast Flow（使用過度表現之ALD4p）進行分離，以反向吸附純化的蛋白質樣品，正向沖提蛋白質。配合不同pH值和流速，找出並測試是否對樣品流出的前後順序有關及其分離的條件。

關鍵字

純化 ； 逆沖提法 ； 吸附性管柱

並列摘要

Plasmids carrying YNR029Cp or ALD4p gene were transformed into E.coli BL21(DE3), and the individual protein was then expressed and purified. Q-Sepharose Fast Flow, Hydroxylapatite (for purified YNR029Cp), Phenyl Sepharose 6 Fast Flow, Blue Sepharose 6 Fast Flow (for over-expressed ALD4p) were employed in order to reversely adsorb the purified protein sample, followed by forward elution of the protein. With the different pH and flow rate, we tried to identify and test the relationship to the order of elution and the separation conditions.

並列關鍵字

purifie ； Back Flush ； Adsorption Column ； Chromatography

參考文獻

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