Comprehensive Analytical Comparison of Strategies Used for Small Molecule Aptamer Evaluation

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• 作者：Maureen McKeague ; Annalisa De Girolamo ; Stefania Valenzano ; Michelangelo Pascale ; Annamaria Ruscito ; Ranganathan Velu ; Nadine R. Frost ; Kayla Hill ; McKenzie Smith ; Erin M. McConnell ; Maria C. DeRosa
• 刊名：Analytical Chemistry
• 出版年：2015
• 出版时间：September 1, 2015
• 年：2015
• 卷：87
• 期：17
• 页码：8608-8612
• 全文大小：259K
• ISSN：1520-6882

文摘

Nucleic acid aptamers are versatile molecular recognition agents that bind to their targets with high selectivity and affinity. The past few years have seen a dramatic increase in aptamer development and interest for diagnostic and therapeutic applications. As the applications for aptamers expand, the need for a more standardized, stringent, and informative characterization and validation methodology increases. Here we performed a comprehensive analysis of a panel of conventional affinity binding assays using a suite of aptamers for the small molecule target ochratoxin A (OTA). Our results highlight inconsistency between conventional affinity assays and the need for multiple characterization strategies. To mitigate some of the challenges revealed in our head-to-head comparison of aptamer binding assays, we further developed and evaluated a set of novel strategies that facilitate efficient screening and characterization of aptamers in solution. Finally, we provide a workflow that permits rapid and robust screening, characterization, and functional verification of aptamers thus improving their development and integration into novel applications.