Purification of a highly modified RNA-aptamer - Effect of complete denaturation during chromatography on product recovery and specific activity

被引:11
作者
Bridonneau, P
Bunch, S
Tengler, R
Hill, K
Carter, J
Pieken, W
Tinnermeier, D
Lehrman, R
Drolet, DW
机构
[1] NeXstar Pharmaceut Inc, Boulder, CO 80301 USA
[2] Proligo LLC, Boulder, CO 80301 USA
[3] Inhale Therapeut Syst, San Carlos, CA 94070 USA
来源
JOURNAL OF CHROMATOGRAPHY B | 1999年 / 726卷 / 1-2期
关键词
RNA-aptamers;
D O I
10.1016/S0378-4347(99)00037-7
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
To evaluate RNA-aptamers as potential drug candidates, efficient and scaleable purification protocols are needed. Because aptamers are highly structured and rigid molecules, denaturation during the purification process is a critical aspect to obtain a pure and active product. A two-step chromatographic procedure was developed to purify a synthetic anti-VEGF aptamer at the preparative scale. A reversed-phase chromatographic step was optimized with a highly hydrophobic ion pairing reagent, followed by ion-exchange chromatography in which heat and a chaotropic salt were used. Because of the presence of 2'-modified ribose, denaturation conditions had to be optimized in both chromatographic steps to achieve a fully active molecule. (C) 1999 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:237 / 247
页数:11
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