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Multiparameter Particle Display (MPPD): A Quantitative Screening Method for the Discovery of Highly Specific Aptamers
Author(s) -
Wang Jinpeng,
Yu Jingwen,
Yang Qin,
McDermott John,
Scott Alexander,
Vukovich Matthew,
Lagrois Remy,
Gong Qiang,
Greenleaf William,
Eisenstein Michael,
Ferguson B. Scott,
Soh H. Tom
Publication year - 2017
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201608880
Subject(s) - aptamer , computational biology , cell sorting , chemistry , sorting , monoclonal antibody , systematic evolution of ligands by exponential enrichment , selection (genetic algorithm) , selex aptamer technique , microbiology and biotechnology , biology , biochemistry , antibody , computer science , cell , gene , genetics , rna , artificial intelligence , programming language
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 10 7 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.