{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T11:47:05Z","timestamp":1791978425813,"version":"4.4.0"},"reference-count":38,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2023,6,25]],"date-time":"2023-06-25T00:00:00Z","timestamp":1687651200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001711","name":"Swiss National Science Foundation","doi-asserted-by":"publisher","award":["4078P0_198281"],"award-info":[{"award-number":["4078P0_198281"]}],"id":[{"id":"10.13039\/501100001711","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>As the COVID-19 pandemic progresses, new variants of SARS-CoV-2 continue to emerge. This underscores the need to develop optimized tools to study such variants, along with new coronaviruses that may arise in the future. Such tools will also be instrumental in the development of new antiviral drugs. Here, we introduce microscale thermophoresis (MST) as a reliable and versatile tool for coronavirus research, which we demonstrate through three different applications described in this report: (1) binding of the SARS-CoV-2 spike receptor binding domain (RBD) to peptides as a strategy to prevent virus entry, (2) binding of the RBD to the viral receptor ACE2, and (3) binding of the RBD to ACE2 in complex with the amino acid transporter SLC6A20\/SIT1 or its allelic variant rs61731475 (p.Ile529Val). Our results demonstrate that MST is a highly precise approach to studying protein\u2013protein and\/or protein\u2013ligand interactions in coronavirus research, making it an ideal tool for studying viral variants and developing antiviral agents. Moreover, as shown in our results, a unique advantage of the MST assay over other available binding assays is the ability to measure interactions with membrane proteins in their near-native plasma membrane environment.<\/jats:p>","DOI":"10.3390\/v15071432","type":"journal-article","created":{"date-parts":[[2023,6,26]],"date-time":"2023-06-26T02:05:02Z","timestamp":1687745102000},"page":"1432","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Applications of the Microscale Thermophoresis Binding Assay in COVID-19 Research"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-9752-2989","authenticated-orcid":false,"given":"Damian T.","family":"Nydegger","sequence":"first","affiliation":[{"name":"Department of Nephrology and Hypertension, Inselspital, University of Bern, Kinderklinik, Freiburgstrasse 15, 3010 Bern, Switzerland"},{"name":"Department of Biomedical Research, Inselspital, University of Bern, Kinderklinik, Freiburgstrasse 15, 3010 Bern, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9951-1390","authenticated-orcid":false,"given":"Jonai","family":"Pujol-Gim\u00e9nez","sequence":"additional","affiliation":[{"name":"Department of Nephrology and Hypertension, Inselspital, University of Bern, Kinderklinik, Freiburgstrasse 15, 3010 Bern, Switzerland"},{"name":"Department of Biomedical Research, Inselspital, University of Bern, Kinderklinik, Freiburgstrasse 15, 3010 Bern, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3795-2876","authenticated-orcid":false,"given":"Palanivel","family":"Kandasamy","sequence":"additional","affiliation":[{"name":"Department of Nephrology and Hypertension, Inselspital, University of Bern, Kinderklinik, Freiburgstrasse 15, 3010 Bern, Switzerland"},{"name":"Department of Biomedical Research, Inselspital, University of Bern, Kinderklinik, Freiburgstrasse 15, 3010 Bern, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1548-6387","authenticated-orcid":false,"given":"Bruno","family":"Vogt","sequence":"additional","affiliation":[{"name":"Department of Nephrology and Hypertension, Inselspital, University of Bern, Kinderklinik, Freiburgstrasse 15, 3010 Bern, Switzerland"},{"name":"Department of Biomedical Research, Inselspital, University of Bern, Kinderklinik, Freiburgstrasse 15, 3010 Bern, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1946-027X","authenticated-orcid":false,"given":"Matthias A.","family":"Hediger","sequence":"additional","affiliation":[{"name":"Department of Nephrology and Hypertension, Inselspital, University of Bern, Kinderklinik, Freiburgstrasse 15, 3010 Bern, Switzerland"},{"name":"Department of Biomedical Research, Inselspital, University of Bern, Kinderklinik, Freiburgstrasse 15, 3010 Bern, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,6,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/978-1-0716-0900-2_1","article-title":"Coronaviruses: An Updated Overview of Their Replication and Pathogenesis","volume":"Volume 2203","author":"Wang","year":"2020","journal-title":"Methods in Molecular Biology"},{"key":"ref_2","unstructured":"Maier, H.J., and Bickerton, E. (2020). Methods in Molecular Biology, Humana."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Chilamakuri, R., and Agarwal, S. (2021). COVID-19: Characteristics and therapeutics. Cells, 10.","DOI":"10.3390\/cells10020206"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Sharma, A., Farouk, I.A., and Lal, S.K. (2021). COVID-19: A Review on the Novel Coronavirus Disease. Viruses, 13.","DOI":"10.3390\/v13020202"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Khan, M., Adil, S.F., Alkhathlan, H.Z., Tahir, M.N., Saif, S., Khan, M., and Khan, S.T. (2021). COVID-19: A Global Challenge with Old History, Epidemiology and Progress So Far. Molecules, 26.","DOI":"10.3390\/molecules26010039"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"101205","DOI":"10.1016\/j.arr.2020.101205","article-title":"Aging in COVID-19: Vulnerability, immunity and intervention","volume":"65","author":"Chen","year":"2021","journal-title":"Ageing Res. Rev."},{"key":"ref_7","unstructured":"WHO Coronavirus (COVID-19) Dashboard (2023, April 07). WHO Coronavirus (COVID-19) Dashboard With Vaccination Data, Available online: https:\/\/covid19.who.int\/."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1016\/j.ymeth.2012.12.005","article-title":"Microscale thermophoresis quantifies biomolecular interactions under previously challenging conditions","volume":"59","author":"Seidel","year":"2023","journal-title":"Methods"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"042001","DOI":"10.1088\/2050-6120\/ac82a6","article-title":"Microscale thermophoresis as a powerful growing analytical technique for the investigation of biomolecular interaction and the determination of binding parameters","volume":"10","author":"Khan","year":"2022","journal-title":"Methods Appl. Fluoresc."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Magnez, R., Bailly, C., and Thuru, X. (2022). Microscale Thermophoresis as a Tool to Study Protein Interactions and Their Implication in Human Diseases. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23147672"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"342","DOI":"10.1089\/adt.2011.0380","article-title":"Molecular interaction studies using microscale thermophoresis","volume":"9","author":"Wienken","year":"2011","journal-title":"Assay Drug Dev. Technol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.ymeth.2018.02.003","article-title":"Thermophoresis for characterizing biomolecular interaction","volume":"146","author":"Asmari","year":"2018","journal-title":"Methods"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1016\/j.molstruc.2014.03.009","article-title":"MicroScale Thermophoresis: Interaction analysis and beyond","volume":"1077","author":"Wanner","year":"2014","journal-title":"J. Mol. Struct."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"5357","DOI":"10.1038\/s41596-021-00619-w","article-title":"The GFP thermal shift assay for screening ligand and lipid interactions to solute carrier transporters","volume":"16","author":"Chatzikyriakidou","year":"2021","journal-title":"Nat. Protoc."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1007\/s00216-003-2101-0","article-title":"Present and future of surface plasmon resonance biosensors","volume":"377","author":"Homola","year":"2003","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1002\/jmr.1167","article-title":"Applications of isothermal titration calorimetry in pure and applied research-survey of the literature from 2010","volume":"25","author":"Ghai","year":"2012","journal-title":"J. Mol. Recognit."},{"key":"ref_17","first-page":"1020","article-title":"Biochimica et Biophysica Acta Global analysis of riboswitches by small-angle X-ray scattering and calorimetry","volume":"1839","author":"Zhang","year":"2014","journal-title":"BBA-Gene Regul. Mech."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"895","DOI":"10.1002\/cphc.201701253","article-title":"Investigating Protein\u2014Ligand Interactions by Solution Nuclear Magnetic Resonance Spectroscopy","volume":"19","author":"Becker","year":"2018","journal-title":"ChemPhysChem"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.vascn.2018.03.004","article-title":"Establishment of a novel microscale thermophoresis ligand-binding assay for characterization of SLC solute carriers using oligopeptide transporter PepT1 (SLC15 family) as a model system","volume":"92","author":"Hediger","year":"2018","journal-title":"J. Pharmacol. Toxicol. Methods"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e3574","DOI":"10.21769\/BioProtoc.3574","article-title":"Measurement of Protein-Protein Interactions through Microscale Thermophoresis (MST)","volume":"10","author":"Romain","year":"2020","journal-title":"Bio-Protocol"},{"key":"ref_21","first-page":"301","article-title":"Microscale Thermophoresis Quantifies Biomolecular Interactions under Previously Challenging Conditions","volume":"59","author":"Dijkman","year":"2014","journal-title":"Methods"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1016\/j.ab.2007.10.034","article-title":"Site-directed mutagenesis by combination of homologous recombination and DpnI digestion of the plasmid template in Escherichia coli","volume":"373","author":"Li","year":"2008","journal-title":"Anal. Biochem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1444","DOI":"10.1126\/science.abb2762","article-title":"Structural basis for the recognition of SARS-CoV-2 by full-length human ACE2","volume":"367","author":"Yan","year":"2020","journal-title":"Science"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1038\/s41586-020-2179-y","article-title":"Structural basis of receptor recognition by SARS-CoV-2","volume":"581","author":"Shang","year":"2020","journal-title":"Nature"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/B978-0-12-407870-3.00006-8","article-title":"The ARC Channel\u2014An Endogenous Store-Independent Orai Channel","volume":"71","author":"Thompson","year":"2013","journal-title":"Curr. Top. Membr."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1038\/s41594-022-00899-0","article-title":"Structure of a volume-regulated heteromeric LRRC8A\/C channel","volume":"30","author":"Rutz","year":"2022","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"230","DOI":"10.3389\/fnmol.2019.00230","article-title":"Oligomeric Receptor Complexes and Their Allosteric Receptor-Receptor Interactions in the Plasma Membrane Represent a New Biological Principle for Integration of Signals in the CNS","volume":"12","author":"Fuxe","year":"2019","journal-title":"Front. Mol. Neurosci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1515\/bmc-2021-0017","article-title":"SIT1 transporter as a potential novel target in treatment of COVID-19","volume":"12","author":"Semiz","year":"2021","journal-title":"Biomol. Concepts"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13059-021-02454-4","article-title":"Integrative approach identifies SLC6A20 and CXCR6 as putative causal genes for the COVID-19 GWAS signal in the 3p21.31 locus","volume":"22","author":"Kasela","year":"2021","journal-title":"Genome Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1350","DOI":"10.1111\/dme.12528","article-title":"A genetic variant in SLC6A20 is associated with Type 2 diabetes in white-European and Chinese populations","volume":"31","author":"Ling","year":"2014","journal-title":"Diabet. Med."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Severe Covid-19 GWAS Group (2020). Genomewide Association Study of Severe COVID-19 with Respiratory Failure. N. Engl. J. Med., 383, 1522\u20131534.","DOI":"10.1056\/NEJMoa2020283"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Azzar\u00e0, A., Cassano, I., Paccagnella, E., Tirindelli, M.C., Nobile, C., Schittone, V., Lintas, C., Sacco, R., and Gurrieri, F. (2022). Genetic variants determine intrafamilial variability of SARS-CoV-2 clinical outcomes in 19 Italian families. PLoS ONE, 17.","DOI":"10.1371\/journal.pone.0275988"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1038\/s41421-022-00488-x","article-title":"Structures of ACE2\u2013SIT1 recognized by Omicron variants of SARS-CoV-2","volume":"8","author":"Shen","year":"2022","journal-title":"Cell Discov."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"zqab027","DOI":"10.1093\/function\/zqab027","article-title":"B0AT1 Amino Acid Transporter Complexed With SARS-CoV-2 Receptor ACE2 Forms a Heterodimer Functional Unit: In Situ Conformation Using Radiation Inactivation Analysis","volume":"2","author":"Stevens","year":"2021","journal-title":"Function"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"591","DOI":"10.1002\/iub.210","article-title":"The role of the neutral amino acid transporter B0AT1 (SLC6A19) in Hartnup disorder and protein nutrition","volume":"61","year":"2009","journal-title":"IUBMB Life"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"e00692-18","DOI":"10.1128\/JVI.00692-18","article-title":"Dimerization of Coronavirus nsp9 with Diverse Modes Enhances Its Nucleic Acid Binding Affinity","volume":"92","author":"Zeng","year":"2018","journal-title":"J. Virol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e00893-18","DOI":"10.1128\/JVI.00893-18","article-title":"Structural and Biochemical Characterization of Endoribonuclease Nsp15 Encoded by Middle East Respiratory Syndrome Coronavirus","volume":"92","author":"Zhang","year":"2018","journal-title":"J. Virol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Shahhamzehei, N., Abdelfatah, S., and Efferth, T. (2022). In Silico and In Vitro Identification of Pan-Coronaviral Main Protease Inhibitors from a Large Natural Product Library. Pharmaceuticals, 15.","DOI":"10.3390\/ph15030308"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/15\/7\/1432\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:00:00Z","timestamp":1760126400000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/15\/7\/1432"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,6,25]]},"references-count":38,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2023,7]]}},"alternative-id":["v15071432"],"URL":"https:\/\/doi.org\/10.3390\/v15071432","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,6,25]]}}}