改进葡聚糖接枝技术制备高性能层析介质

doi:10.16085/j.issn.1000-6613.2018-1978

化工进展 ›› 2019, Vol. 38 ›› Issue (06): 2658-2664.DOI: 10.16085/j.issn.1000-6613.2018-1978

改进葡聚糖接枝技术制备高性能层析介质

张强¹(),操文军¹,夏海锋^1,²(),丁重阳^1,²()

1. 江南大学生物工程学院，工业生物技术教育部重点实验室，江苏无锡 214122
2. 江南大学粮食发酵工艺与;技术国家工程实验室，江苏无锡 214122

收稿日期:2018-09-30 出版日期:2019-06-05 发布日期:2019-06-05
通讯作者: 夏海锋,丁重阳
作者简介:张强（1992—），男，硕士研究生，研究方向为离子交换色谱。E-mail：<email>6160205031@vip.jiangnan.edu.cn</email>。
基金资助:
国家重点研发计划(2018YFC1603202，2018YFC1603200)

Preparation of high performance chromatography media by improved dextran grafting technology

Qiang ZHANG¹(),Wenjun CAO¹,Haifeng XIA^1,²(),Chongyang DING^1,²()

1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China
2. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, Jiangsu, China

Received:2018-09-30 Online:2019-06-05 Published:2019-06-05
Contact: Haifeng XIA,Chongyang DING

摘要/Abstract

摘要：

利用改进的葡聚糖接枝技术，在以环氧氯丙烷为交联剂交联琼脂糖微球骨架的过程中加入葡聚糖溶液，在交联的同时接枝葡聚糖制得葡聚糖接枝型琼脂糖微球Rigose-Dex，再与盐酸2-氯三乙胺（DEAE）反应，获得葡聚糖接枝型高载量弱阴离子交换介质Rigose-Dex DEAE。以牛血清白蛋白（BSA）为模型蛋白，以商品化介质DEAE Sepharose 6FF为对照，系统研究了该葡聚糖接枝型Rigose-Dex DEAE的蛋白吸附性能，并进行了物理性能研究。结果表明，改进后葡聚糖接枝技术的最高接枝量为24.5mg/mL。自制Rigose-Dex DEAE可耐受700cm/h 的线性流速，对BSA的动态饱和载量为127.6mg/mL, 为商品介质DEAE Sepharose 6FF 载量的212%；具有在高流速下快速结合蛋白的能力，上样蛋白溶液在层析柱中停留2min即可基本达到饱和动态载量；重复使用性能好，经120 次在线清洗后，Rigose-Dex DEAE介质的动态载量为原始载量的90.4%。

关键词: 离子交换, 葡聚糖, 接枝, 介质, 吸附

Abstract:

With the improved dextran grafting technique on agarose beads, dextran solution was added during the process of cross-linking with epichlorohydrin as crosslinker,and the dextran-grafted agarose was named Rigose-Dex. Then Rigose-Dex was coupled with 2-chloro-N,N-diethylethylamine hydrochloride(DEAE) to obtain the weak anion exchanger which named Rigose-Dex DEAE. Protein adsorption properties of Rigose-Dex DEAE were studied systematically using bovine serum albumin (BSA) as the model protein and physical properties were studied, which compared with commercial product DEAE Sepharose 6FF. The results showed that the highest grafting amount of the modified dextran grafting technique was 24.5mg/mL. Rigose-Dex DEAE can tolerate linear flow rate of 700cm/h.The dynamic capacity of BSA of Rigose-Dex DEAE medium was 127.6mg/mL even at the retention time of 2 minutes in the chromatography column, which was 212% of that of DEAE Sepharose 6FF. The Rigose-Dex DEAE showed excellent stability with 90.4% capacity remaining after 120 cycles of on-line cleaning.

Key words: ion exchange, dextran, graft, media, adsorption

中图分类号:

TQ033

张强, 操文军, 夏海锋, 丁重阳. 改进葡聚糖接枝技术制备高性能层析介质[J]. 化工进展, 2019, 38(06): 2658-2664.

Qiang ZHANG, Wenjun CAO, Haifeng XIA, Chongyang DING. Preparation of high performance chromatography media by improved dextran grafting technology[J]. Chemical Industry and Engineering Progress, 2019, 38(06): 2658-2664.

图/表 9

图1 葡聚糖接枝型高强度Rigose-Dex介质的制备过程示意图

图2 层析介质葡聚糖接枝量的测定

图3 Rigose-Dex介质扫描电镜照片

图4 3种介质粒径分布图

图5 Rigose-Dex DEAE与DEAE Sepharose 6FF反压-流速关系图

图6 BSA在两种层析介质的等温吸附曲线

图7 两种层析介质的穿透曲线

图8 重复使用性能测试图

图9 不同停留时间下介质的动态载量图

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改进葡聚糖接枝技术制备高性能层析介质

Preparation of high performance chromatography media by improved dextran grafting technology

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