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).
pp. 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 |
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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: | 19 Jan 2023 01:06 |
DOI: | 10.1021/acs.analchem.6b00813 |
Open Access URL: | https://www.research.manchester.ac.uk/portal/en/pu... |
Related URLs: | |
URI: | https://livrepository.liverpool.ac.uk/id/eprint/3031268 |
Available Versions of this Item
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Label-Free Surface Enhanced Raman Scattering Approach for High-Throughput Screening of Biocatalysts. (deposited 04 May 2016 07:45)
- Label-Free Surface Enhanced Raman Scattering Approach for High-Throughput Screening of Biocatalysts. (deposited 15 Jan 2019 09:48) [Currently Displayed]