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



Westley, C, Xu, Y ORCID: 0000-0003-3228-5111, Carnell, AJ ORCID: 0000-0002-6353-403X, Turner, NJ and Goodacre, R ORCID: 0000-0003-2230-645X
(2016) Label-Free Surface Enhanced Raman Scattering Approach for High-Throughput Screening of Biocatalysts. Analytical Chemistry, 88 (11). 5898 - 5903.

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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.

Item Type: Article
Uncontrolled Keywords: high-performance liquid chromatography, peptides and proteins, drug discovery, Raman spectroscopy, screening assays
Depositing User: Symplectic Admin
Date Deposited: 15 Jan 2019 09:48
Last Modified: 27 Sep 2021 19:15
DOI: 10.1021/acs.analchem.6b00813
Open Access URL: https://www.research.manchester.ac.uk/portal/en/pu...
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URI: https://livrepository.liverpool.ac.uk/id/eprint/3031268

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