A novel cell-based binding assay system reconstituting interaction between SARS-CoV S protein and its cellular receptor

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dc.contributor.authorChou, Chih-Fong
dc.contributor.authorShen, Shuo
dc.contributor.authorTan, Yee-Joo
dc.contributor.authorFielding, Burtram C.
dc.contributor.authorTan, Timothy H.P.
dc.contributor.authorFu, Jianlin
dc.contributor.authorXu, Qiurong
dc.contributor.authorLim, Seng Gee
dc.contributor.authorHong, Wanjin
dc.date.accessioned2013-10-17T20:45:30Z
dc.date.available2013-10-17T20:45:30Z
dc.date.issued2005
dc.description.abstractSevere acute respiratory syndrome (SARS), a life-threatening disease, is caused by the newly identified virus SARS coronavirus (SARSCoV). In order to study the spike (S) protein of this highly contagious virus, we established a clonal cell-line, CHO-SG, from the Chinese hamster ovary cells that stably expresses C-terminally EGFP-tagged SARS-CoV S protein (S-EGFP). The ectodomain of the S glycoprotein is localized on the surface of CHO-SG cells with N-acetyl-glucosamine-terminated carbohydrate structure. CHO-SG cells associated tightly with Vero E6 cells, a SARS-CoV receptor (ACE2) expressing cell-line, and the interaction remained stable under highly stringent condition (1MNaCl). This interaction could be blocked by either the serum from a SARS convalescent patient or a goat anti-ACE2 antibody, indicating that the interaction is specific. A binding epitope with lesser degree of glycosylation and native conformation was localized by using rabbit anti-sera raised against five denatured recombinant S protein fragments expressed in Escherichia coli. One of the sera obtained from the fragment encompassing amino acids 48-358 significantly blocked the interaction between CHO-SG and Vero E6 cells. The region is useful for studying neutralizing antibodies in future vaccine development. This paper describes an easy and safe cell-based assay suitable for studying the binding between SARS-CoV S protein and its receptor.en_US
dc.description.accreditationWeb of Scienceen_US
dc.identifier.citationChou, C-F, et al. (2005). A novel cell-based binding assay system reconstituting interaction between SARS-CoV S protein and its cellular receptor. Journal of Virological Methods, 123(1):41-8en_US
dc.identifier.issn0166-0934
dc.identifier.urihttp://hdl.handle.net/10566/769
dc.language.isoenen_US
dc.privacy.showsubmitterfalse
dc.publisherElsevieren_US
dc.rightsCopyright Elsevier
dc.source.urihttp://dx.doi.org/10.1016/j.jviromet.2004.09.008
dc.status.ispeerreviewedtrue
dc.subjectSARS-CoVen_US
dc.subjectEctodomainen_US
dc.subjectCHOen_US
dc.subjectEGFP-taggeden_US
dc.subjectACE2en_US
dc.subjectCell-based assayen_US
dc.titleA novel cell-based binding assay system reconstituting interaction between SARS-CoV S protein and its cellular receptoren_US
dc.typeArticleen_US

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