Improving aptamer selection efficiency through volume dilution, magnetic concentration, and continuous washing in microfluidic channels.
Improving aptamer selection efficiency through volume dilution, magnetic concentration, and continuous washing in microfluidic channels.
复制标题
DOI：
10.1021/ac201269f
复制
发表时间：
2011-09-01
影响因子：
7.4
通讯作者：
Soh, H. Tom
中科院分区：
文献类型：
--
作者：
Oh, Seung Soo;Ahmad, Kareem M.;Cho, Minseon;Kim, Seon;Xiao, Yi;Soh, H. Tom
具有高亲和力的核酸适体的产生通常需要耗时的、反复的结合、分离和扩增过程。因此，开发一种有效的选择策略可以快速、经济且可重复地生成这些高质量的适体将是有益的。为了实现这一目标，我们开发了一种能够有效生成解离速率较慢的 DNA 适体的方法。这种方法称为 VDC-MSELEX，将体积稀释挑战过程与微流体分离相结合，以进行磁珠辅助适体选择。该方法通过应用高度严格的选择条件来提高适体选择效率：它回收悬浮在大体积（> 50 mL）中的少量（< 106）磁珠，并将它们浓缩到微流体室（8 μL）中，以最小的损失进行连续清洗。我们以链霉亲和素（SA）为靶标进行了三轮VDC-MSELEX，获得了具有低纳摩尔亲和力、特异性结合SA蛋白的新DNA适体序列。
The generation of nucleic acid aptamers with high affinity typically entails a time-consuming, iterative process of binding, separation, and amplification. It would therefore be beneficial to develop an efficient selection strategy that can generate these high quality aptamers rapidly, economically, and reproducibly. Toward this goal, we have developed a method that efficiently generates DNA aptamers with slow off-rates. This methodology, called VDC-MSELEX, pairs the volume dilution challenge process with microfluidic separation for magnetic bead-assisted aptamer selection. This method offers improved aptamer selection efficiencies through the application of highly stringent selection conditions: it retrieves a small number (< 106) of magnetic beads suspended in a large volume (> 50 mL) and concentrates them into a microfluidic chamber (8 μL) with minimal loss for continuous washing. We performed three rounds of the VDC-MSELEX using streptavidin (SA) as the target, and obtained new DNA aptamer sequences with low nanomolar affinity that specifically bind to the SA proteins.
登录
查看更多内容
影响因子：
3.7
作者：
Gold L;Ayers D;Bertino J;Bock C;Bock A;Brody EN;Carter J;Dalby AB;Eaton BE;Fitzwater T;Flather D;Forbes A;Foreman T;Fowler C;Gawande B;Goss M;Gunn M;Gupta S;Halladay D;Heil J;Heilig J;Hicke B;Husar G;Janjic N;Jarvis T;Jennings S;Katilius E;Keeney TR;Kim N;Koch TH;Kraemer S;Kroiss L;Le N;Levine D;Lindsey W;Lollo B;Mayfield W;Mehan M;Mehler R;Nelson SK;Nelson M;Nieuwlandt D;Nikrad M;Ochsner U;Ostroff RM;Otis M;Parker T;Pietrasiewicz S;Resnicow DI;Rohloff J;Sanders G;Sattin S;Schneider D;Singer B;Stanton M;Sterkel A;Stewart A;Stratford S;Vaught JD;Vrkljan M;Walker JJ;Watrobka M;Waugh S;Weiss A;Wilcox SK;Wolfson A;Wolk SK;Zhang C;Zichi D
通讯作者：
Zichi D
链霉亲和素高亲和力 DNA 适体的体外筛选
影响因子：
3.7
作者：
Wang, Chenglong;Yang, Guang;Ding, Hongmei
通讯作者：
Ding, Hongmei
影响因子：
15
作者：
Luchansky, SJ;Nolan, SJ;Baranger, AM
通讯作者：
Baranger, AM
DOI：
10.3390/molecules15010001
发表时间：
2009-12-24
期刊：
Molecules (Basel, Switzerland)
影响因子：
--
作者：
Avci-Adali M;Paul A;Wilhelm N;Ziemer G;Wendel HP
通讯作者：
Wendel HP
DOI：
10.1073/pnas.1009331107
发表时间：
2010-08-31
影响因子：
11.1
作者：
Cho, Minseon;Xiao, Yi;Soh, H. Tom
通讯作者：
Soh, H. Tom