Multiparameter Particle Display (MPPD): A Quantitative Screening Method for the Discovery of Highly Specific Aptamers
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- 作者：Dr. Jinpeng Wang ; Jingwen Yu ; Dr. Qin Yang ; John McDermott ; Alexander Scott ; Matthew Vukovich ; Remy Lagrois ; Dr. Qiang Gong ; Prof. William Greenleaf ; Dr. Michael Eisenstein ; Dr. B. Scott Ferguson and Prof. H. Tom Soh
- 刊名：Angewandte Chemie
- 出版年：2017
- 出版时间：January 16, 2017
- 年：2017
- 卷：129
- 期：3
- 页码：762-765
- 全文大小：2540K
- ISSN：1521-3757
文摘
Aptamers are a promising class of affinity reagents because they are chemically synthesized, thus making them highly reproducible and distributable as sequence information rather than a physical entity. Although many high-quality aptamers have been previously reported, it is difficult to routinely generate aptamers that possess both high affinity and specificity. One of the reasons is that conventional aptamer selection can only be performed either for affinity (positive selection) or for specificity (negative selection), but not both simultaneously. In this work, we harness the capacity of fluorescence activated cell sorting (FACS) for multicolor sorting to simultaneously screen for affinity and specificity at a throughput of 107 aptamers per hour. As a proof of principle, we generated DNA aptamers that exhibit picomolar to low nanomolar affinity in human serum for three diverse proteins, and show that these aptamers are capable of outperforming high-quality monoclonal antibodies in a standard ELISA detection assay.