Label-Free Surface Enhanced Raman Scattering Approach for High-Throughput Screening of Biocatalysts

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• 作者：Chloe Westley ; Yun Xu ; Andrew J. Carnell ; Nicholas J. Turner ; Royston Goodacre
• 刊名：Analytical Chemistry
• 出版年：2016
• 出版时间：June 7, 2016
• 年：2016
• 卷：88
• 期：11
• 页码：5898-5903
• 全文大小：403K
• 年卷期：0
• ISSN：1520-6882

文摘

Biocatalyst discovery and directed evolution are central to many pharmaceutical research programs, yet the lack of robust high-throughput screening methods for large libraries of enzyme variants generated (typically 10⁶–10⁸) has hampered progress and slowed enzyme optimization. We have developed a label-free generally applicable approach based on Raman spectroscopy which results in significant reductions in acquisition times (>30-fold). Surface enhanced Raman scattering (SERS) is employed to monitor the enzyme-catalyzed conversion by xanthine oxidase of hypoxanthine to xanthine to uric acid. This approach measures the substrates and products directly and does not require chromogenic substrates or lengthy chromatography, was successfully benchmarked against HPLC, and shows high levels of accuracy and reproducibility. Furthermore, we demonstrate that this SERS approach has utility in monitoring enzyme inhibition illustrating additional medical significance to this high-throughput screening method.