Screening Platform toward New Anti-HIV Aptamers Set on Molecular Docking and Fluorescence Quenching Techniques

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• 作者：Giorgia Oliviero ; Mariano Stornaiuolo ; Valentina D’Atri ; Fabrizia Nici ; Ali Munaim Yousif ; Stefano D’Errico ; Gennaro Piccialli ; Luciano Mayol ; Ettore Novellino ; Luciana Marinelli ; Paolo Grieco ; Alfonso Carotenuto ; Sam Noppen ; Sandra Liekens ; Jan Balzarini ; Nicola Borbone
• 刊名：Analytical Chemistry
• 出版年：2016
• 出版时间：February 16, 2016
• 年：2016
• 卷：88
• 期：4
• 页码：2327-2334
• 全文大小：448K
• ISSN：1520-6882

文摘

By using a new rapid screening platform set on molecular docking simulations and fluorescence quenching techniques, three new anti-HIV aptamers targeting the viral surface glycoprotein 120 (gp120) were selected, synthesized, and assayed. The use of the short synthetic fluorescent peptide V35-Fluo mimicking the V3 loop of gp120, as the molecular target for fluorescence-quenching binding affinity studies, allowed one to measure the binding affinities of the new aptamers for the HIV-1 gp120 without the need to obtain and purify the full recombinant gp120 protein. The almost perfect correspondence between the calculated K_d and the experimental EC₅₀ on HIV-infected cells confirmed the reliability of the platform as an alternative to the existing methods for aptamer selection and measuring of aptamer–protein equilibria.