Accelerating the Throughput of Affinity Mass Spectrometry-Based Ligand Screening toward a G Protein-Coupled Receptor
Open accessAbstract
Affinity mass spectrometry (MS) enables rapid screening of compound mixtures for ligands bound to a specific protein target, yet its current throughput is limited to individually assay pools of 400–2000 compounds. Typical affinity MS screens implemented in pharmaceutical industry laboratories identify putative ligands based on qualitative analysis of compound binding to the target whereas no quantitative information is acquired to discriminate high- and low-affinity ligands in the screening phase. Furthermore, these screens require purification of a stabilized form of the protein target, which poses a great challenge for membrane receptor targets. Here, we describe a new, potentially general affinity MS strategy that allows screening of 20,000 compounds in one pool for highly efficient ligand discovery toward a G protein-coupled receptor (GPCR) target. Quantitative measurement of compound binding to the receptor enables high-affinity ligand selection using both the purified receptor and receptor-embedded cell membranes. This high-throughput, label-free and quantitative affinity MS screen resulted in discovery of three new antagonists of the A 2A adenosine receptor.
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AS-MS can screen large numbers of compounds, and up to 20,000 compounds in one sample have been analyzed so far.
“This high-throughput method can be used to screen large numbers of compounds,72,73 up to 20 000 compounds in one sample have been analyzed so far.74”
From Connecting metabolome and phenotype: recent advances in functional metabolomics tools for the identification of bioactive natural products (2024) · cites this paper as [74]
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- [23]Fu X; Wang Z; Li L; Dong S; Li Z; Jiang Z; Wang Y; Shui W Sci. Rep 2016, 6, 29680. 27403722 PMC4940736
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- Authors
- Lu, Y., Qin, S., Zhang, B., Dai, A., Cai, X., Ma, M., Gao, Z.-G., Yang, D., Stevens, R. C., Jacobson, K. A., Wang, M.-W., & Shui, W.
- Affiliations
- ShanghaiTech University (CN)
- Chinese Academy of Sciences (CN)
- National Institutes of Health (US)
- Corresponding author
- Wang, Ming-Wei
- Shui, Wenqing
- ORCID
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2019-05-16
- Volume
- 91
- Issue
- 13
- Pages
- 8162-8169
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- PMC
- PMC6669887
- OpenAlex
- W2946364931
- Cited by
- 39
- References
- 30
- Citations by year
- 2026: 2 · 2025: 9 · 2024: 5 · 2023: 6 · 2022: 3 · 2021: 4 · 2020: 9 · 2019: 1
- FWCI
- 1.69
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- Locations
- Analytical Chemistry (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- Ministry of Science and Technology of the People's Republic of China (2018YFA0507004, 2018ZX09735-001, 2018ZX09711002-002-005)
- National Institute of Diabetes and Digestive and Kidney Diseases (ZIADK31117)
- National Natural Science Foundation of China (81573479, 81773792)
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolomics and Mass Spectrometry Studies
- Keywords
- ligand screening, G protein-coupled receptors, adenosine A2A receptor, high-throughput screening, cell surface receptors, receptor antagonists, label-free screening
- MeSH terms
- Cell Membrane, Humans, Ligands, Protein Binding, Mass Spectrometry, Receptors, G-Protein-Coupled, High-Throughput Screening Assays, Adenosine A2 Receptor Antagonists
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