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

Chloe Westley, Yun Xu, Andrew J. Carnell, Nicholas Turner, Royston Goodacre

    Research output: Contribution to journalArticlepeer-review

    183 Downloads (Pure)

    Abstract

    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.

    Original languageEnglish
    Pages (from-to)5898-5903
    Number of pages6
    JournalAnalytical Chemistry
    Volume88
    Issue number11
    Early online date30 Apr 2016
    DOIs
    Publication statusPublished - 30 Apr 2016

    Fingerprint

    Dive into the research topics of 'Label-Free Surface Enhanced Raman Scattering Approach for High-Throughput Screening of Biocatalysts'. Together they form a unique fingerprint.

    Cite this