Label-Free Real-Time Microarray Imaging of Cancer Protein–Protein Interactions and Their Inhibition by Small Molecules

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• 作者：Charuksha Walgama ; Zainab H. Al Mubarak ; Bing Zhang ; Mayowa Akinwale ; Anuruddha Pathiranage ; Junpeng Deng ; K. Darrell Berlin ; Doris M. Benbrook ; Sadagopan Krishnan
• 刊名：Analytical Chemistry
• 出版年：2016
• 出版时间：March 15, 2016
• 年：2016
• 卷：88
• 期：6
• 页码：3130-3135
• 全文大小：380K
• ISSN：1520-6882

文摘

A rapid optical microarray imaging approach for anticancer drug screening at specific cancer protein–protein interface targets with binding kinetics and validation by a mass sensor is reported for the first time. Surface plasmon resonance imager (SPRi) demonstrated a 3.5-fold greater specificity for interactions between murine double minute 2 protein (MDM2) and wild-type p53 over a nonspecific p53 mutant in a real-time microfluidic analysis. Significant percentage reflectivity changes (Δ%R) in the SPRi signals and molecular-level mass changes were detected for both the MDM2–p53 interaction and its inhibition by a small-molecule Nutlin-3 drug analogue known for its anticancer property. We additionally demonstrate that synthetic, inexpensive binding domains of interacting cancer proteins are sufficient to screen anticancer drugs by an array-based SPRi technique with excellent specificity and sensitivity. This imaging array, combined with a mass sensor, can be used to study quantitatively any protein–protein interaction and screen for small molecules with binding and potency evaluations.