[1]唐明青,张文豪,侯莹,等.超滤法去除重组腺相关病毒中内毒素的应用[J].华侨大学学报(自然科学版),2017,38(1):64-68.[doi:10.11830/ISSN.1000-5013.201701012]
　TANG Mingqing,ZHANG Wenhao,HOU Ying,et al.Removal of Endotoxin From Recombinant Adeno-AssociatedVirus by Ultrafiltration[J].Journal of Huaqiao University(Natural Science),2017,38(1):64-68.[doi:10.11830/ISSN.1000-5013.201701012]
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超滤法去除重组腺相关病毒中内毒素的应用()
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《华侨大学学报（自然科学版）》[ISSN:1000-5013/CN:35-1079/N]

卷:
第38卷
期数:
2017年第1期
页码:
64-68
栏目:
出版日期:
2017-01-09

文章信息/Info

Title:
Removal of Endotoxin From Recombinant Adeno-AssociatedVirus by Ultrafiltration
文章编号:
1000-5013(2017)01-0064-05
作者:
唐明青1;  张文豪1; 2;  侯莹1;  成志云1; 郭思佳1;  王立振1;  刘慧1;  许瑞安1
1. 华侨大学 生物医学学院, 福建 泉州 362021;2. 福建医科大学 基础医学院, 福建 福州 350108
Author(s):
TANG Mingqing1;  ZHANG Wenhao1; 2;  HOU Ying1;  CHENG Zhiyun1;  GUO Sijia1;  WANG Lizhen1;  LIU Hui1;  XU Ruian1
1. School of Biomedical Sciences, Huaqiao University, Quanzhou 362021, China; 2. School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350108, China
关键词:
重组腺相关病毒;  超滤;  截留分子量;  内毒素;  纯化
Keywords:
recombinant adeno-associated virus;  ultrafiltration;  molecular weights cut off;  endotoxin;  purification
分类号:
Q93
DOI:
10.11830/ISSN.1000-5013.201701012
文献标志码:
A
摘要:
为探讨超滤法在重组腺相关病毒精制中的可行性,采用不同截留分子量的超滤管对重组腺相关病毒样品rAAV9-Kal进行超滤处理.利用鲎试剂盒和qPCR法定量检测截留液和滤过液中的内毒素和病毒含量,评价超滤技术对样品中内毒素的去除效果.结果表明:在乙二胺四乙酸二钠和脱氧胆酸钠的预处理下,30 kD的超滤管对样品的回收率及内毒素去除率分别为0.912 2,0.870 6.因此,在单一超滤条件下,30 kD的超滤管是重组腺相关病毒样品超滤纯化的最佳选择.
Abstract:
To explore the possibility of ultrafiltration technology used in recombinant adeno-associated virus purification, the recombinant adeno-associated virus sample termed rAAV9-Kal was first treated by different molecular weights cut off ultrafiltration devices, then the endotoxin and virus in the retentates and filtrates were assayed with quantitative tachypleus amebocyte lysate regents and real-time PCR respectively, and last the effects of ultrafiltration on endotoxin removal were evaluated. The results showed that the recovery rate and endotoxin removal rate of sample were 0.912 2 and 0.870 6 respectively, under the treatment of EDTA, DOC and 30 kD ultrafiltration devices. Therefore, the 30 kD should be the most suitable ultrafiltration device for sample ultrafiltration and endotoxin removal.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期: 2016-06-29
通信作者: 唐明青(1982-),男,副教授,博士,主要从事AAV基因治疗的研究.E-mail:Tangmingqing2222@163.com
基金项目: 国家自然科学基金资助项目(2012AA020810); 福建省中青年教师教育科研项目(JA14020); 华侨大学科研基金资助项目(14BS111)
更新日期/Last Update: 2017-01-20