Multiparameter Particle Display (MPPD): A Quantitative Screening Method for the Discovery of Highly Specific Aptamers.
Multiparameter Particle Display (MPPD): A Quantitative Screening Method for the Discovery of Highly Specific Aptamers.
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DOI：
10.1002/anie.201608880
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发表时间：
2017-01-16
期刊：
影响因子：
--
通讯作者：
Soh HT
中科院分区：
文献类型：
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作者：
Wang J;Yu J;Yang Q;McDermott J;Scott A;Vukovich M;Lagrois R;Gong Q;Greenleaf W;Eisenstein M;Ferguson BS;Soh HT
适体是一类很有前途的亲和试剂，因为它们是化学合成的，使它们作为序列信息而不是物理实体具有高度的可重复性和可分布性。虽然许多高质量的适体已被报道，但很难常规地产生同时具有高亲和力和特异性的适体。原因之一是因为常规适体选择只能针对亲和力（阳性选择）或特异性（阴性选择）进行，而不能同时进行。在这项工作中，我们利用荧光激活细胞分选（FACS）进行多色分选的能力，以每小时107个适体的通量同时筛选亲和力和特异性。作为原理的证明，我们产生了三种不同蛋白质的DNA适体，这些蛋白质在人血清中表现出皮摩尔至低纳摩尔的亲和力，并表明这些适体能够在标准ELISA检测测定中优于高质量的单克隆抗体。
Aptamers are a promising class of affinity reagents because they are chemically synthesized, 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 because 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 ability of fluorescence activated cell sorting (FACS) to perform multi-color sorting to simultaneously screen for affinity and specificity at a throughput of 107 aptamers per hour. As proof of principle, we generated DNA aptamers for three diverse proteins that exhibit picomolar to low nanomolar affinity in human serum, and show that these aptamers are capable of outperforming high-quality monoclonal antibodies in a standard ELISA detection assay.