文献类型：期刊文献

中文题名：头孢唑啉为配基亲和层析纯化尿激酶

英文题名：Affinity Chromatography of Urokinase Using Cefazoline as a Ligand

作者：曹学君[1];邬行彦[1];朱开昱[1];戴干策[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室

年份：1998

卷号：30

期号：6

起止页码：635

中文期刊名：生物化学与生物物理学报

收录：CSTPCD;;Scopus;CSCD:【CSCD2011_2012】；PubMed;

语种：中文

中文关键词：头孢唑啉;亲和层析;尿激酶;琼脂糖凝胶

外文关键词：Cefazoline; affinity chromatography; urokinase; agarose; epichlorohydrin

摘要：利用头孢类抗生素头孢唑啉为配基，Ｓｅｐｈａｒｏｓｅ４Ｂ为载体，用环氧氯丙烷活化法制得头孢唑啉－Ｓｅｐｈａｒｏｓｅ４Ｂ亲和载体，配基密度为４３μｍｏｌ／ｇ湿胶。吸附尿激酶的最佳条件为ｐＨ６．０和１．０ｍｏｌ／ＬＮａＣｌ；最大吸附量为１１００００ｕ／ｇ湿胶，洗脱条件为含０．５ｍｏｌ／ＬＮａＣｌ，ｐＨ９．０，０．１ｍｏｌ／Ｌ甘氨酸缓冲液。将比活为５００ｕ／ｍｇ的尿激酶粗品进行层析，得到比活为４９３００ｕ／ｍｇ的产品，活性回收率为７８％，纯化倍数为９８．６。
Sepharose 4B based affinity adsorbent was prepared using cefazoline as a ligand, and epichlorohydrin as an activator .The density of the ligand in the affinity adsorbents was about 43 μmol/g wet gel. The optimal adsorption pH for urokinase was 6.0, and the optimal adsorption salt concentration was 1.0 mol/L NaCl. The optimal conditions of elution was 0.1 mol/L glycine buffer, pH 9.0, containing 0.5 mol/L NaCl. A crude urokinase with specific activity 500 u/mg protein was purified on an affinity chromatography column (1 cm×12 cm), yielding a product of specific activity of 49 300 u/mg with 78% recovery and 98.6 fold purification.