Pressure controllable aptamers picking strategy by targets competition
Pressure controllable aptamers picking strategy by targets competition
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目标竞争的压力可控适体挑选策略
DOI：
10.1016/j.cclet.2020.10.018
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发表时间：
2021
影响因子：
9.1
通讯作者：
Feng Qu
中科院分区：
文献类型：
--
作者：
Ge Yang;Chao Zhu;Liping Zhao;Linsen Li;Yuanyu Huang;Yukui Zhang;Feng Qu
选择具有高亲和力和良好特异性的适体需要多轮分离和PCR扩增的交替步骤。在此，我们提出了一种新的高效的适体挑选策略：一轮压力可控选择（OPCS）。OPCS集筛选优势、高效分离、一轮筛选和PCR扩增、同步阴性选择和靶标竞争四种模式于一体。通过调节蛋白质浓度的平衡竞争和引入高浓度的捕食性蛋白质的优势竞争，可以实现筛选压力的可控。在OPCS过程中，两种蛋白质与一个ssDNA文库共孵育，每种蛋白质分别与其有利序列特异性结合，形成蛋白质-ssDNA复合物。同时，一种蛋白质可以提供/承受另一种蛋白质的亲和力和特异性的挑选压力，这消除了彼此弱结合或未结合的序列。两种复合物可方便地分离和收集，同时获得两种蛋白质的适体，具有高亲和力和良好的特异性。这一策略不仅为适体的筛选提供了一种更有效的方法，而且在其他配体或药物的筛选中显示出巨大的潜力。
Selection of aptamers with high affinity and good specificity requires multiple rounds of alternating steps of separation and PCR amplification. Herein, we proposed a novel high-efficiency aptamers picking strategy: One-round pressure controllable selection(OPCS). OPCS integrates four types of screening superiority, high-efficiency separation, one-round selection and PCR amplification, synchronous negative selection and targets competition. The controllable screening pressure can be achieved through two approaches, balanced competition by the regulation of protein concentration, and dominant competition by introducing a predatory protein with high concentration. In OPCS process, two proteins were co-incubated with one ssDNA library, and each protein bound its favorable sequences specifically and formed protein-ssDNA complex respectively. Meanwhile, one protein could supply/suffer the picking pressure of affinity and specificity to/from another, which eliminated weakly bound or unbound sequences for each other. Two complexes could be separated and collected conveniently, and aptamers for two proteins obtained synchronously with high affinity and good specificity. This strategy not only provides a more effective way for aptamers selection, but shows great potential in other ligands or drugs selection.
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DOI：
10.1016/j.chroma.2006.03.101
发表时间：
2006-06
期刊：
Journal of chromatography. A
影响因子：
--
作者：
C. Ravelet;C. Grosset;E. Peyrin
通讯作者：
C. Ravelet;C. Grosset;E. Peyrin
多功能毛细管电泳适配体高效选择的进展
影响因子：
16
作者：
Chao Zhu;Ge Yang;Ghulam Murtaza;Linsen Li;Feng Qu
通讯作者：
Feng Qu
影响因子：
6.2
作者：
S. Lisi;Emmanuelle Fiore;S. Scarano;E. Pascale;Yannik Boehman;F. Ducongé;S. Chierici;M. Minunni;E. Peyrin;C. Ravelet
通讯作者：
S. Lisi;Emmanuelle Fiore;S. Scarano;E. Pascale;Yannik Boehman;F. Ducongé;S. Chierici;M. Minunni;E. Peyrin;C. Ravelet
影响因子：
15
作者：
Mendonsa, SD;Bowser, MT
通讯作者：
Bowser, MT
影响因子：
1.2
作者：
Li-Ping ZHAO;Ge YANG;Xiao-Min ZHANG;Feng QU
通讯作者：
Feng QU