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己糖胺酶的亲和纯化

Affinity purification of hexosaminidases.

作者信息

Sankaranarayanan P, Chandrasekaran S, Puvanakrishnan R, Dhar S C, Gopalakrishnan S, Rangabashyam N

机构信息

Institute of Biochemistry, Madras Medical College, India.

出版信息

J Biochem Biophys Methods. 1987 Dec;15(3-4):207-14. doi: 10.1016/0165-022x(87)90119-9.

Abstract

Hexosaminidases A and B were purified by affinity chromatography from normal gastric mucosa, after preliminary separation of isozymes by anion exchange chromatography. Heparin and mannosamine were coupled to Sepharose 4B and used as affinity matrices and the purified enzymes were found to be homogeneous when analysed by polyacrylamide slab gel electrophoresis. This combination of 2 novel affinity chromatographic procedures is superior to existing methods in that a final yield of over 70% could be achieved. Also, the number of steps required to obtain homogeneity are less in contrast to the conventional methods used previously.

摘要

在通过阴离子交换色谱对同工酶进行初步分离后,从正常胃黏膜中通过亲和色谱法纯化己糖胺酶A和B。将肝素和甘露糖胺偶联到琼脂糖凝胶4B上,并用作亲和基质,通过聚丙烯酰胺平板凝胶电泳分析时,发现纯化后的酶是均一的。这两种新型亲和色谱方法的组合优于现有方法,因为最终产率可达到70%以上。此外,与之前使用的传统方法相比,获得均一性所需的步骤更少。

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