dc.contributor.author | Chou, Chih-Fong | |
dc.contributor.author | Shen, Shuo | |
dc.contributor.author | Tan, Yee-Joo | |
dc.contributor.author | Fielding, Burtram C. | |
dc.contributor.author | Tan, Timothy H.P. | |
dc.contributor.author | Fu, Jianlin | |
dc.contributor.author | Xu, Qiurong | |
dc.contributor.author | Lim, Seng Gee | |
dc.contributor.author | Hong, Wanjin | |
dc.date.accessioned | 2013-10-17T20:45:30Z | |
dc.date.available | 2013-10-17T20:45:30Z | |
dc.date.issued | 2005 | |
dc.identifier.citation | Chou, 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):41-8 | en_US |
dc.identifier.issn | 0166-0934 | |
dc.identifier.uri | http://hdl.handle.net/10566/769 | |
dc.description.abstract | Severe 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.language.iso | en | en_US |
dc.publisher | Elsevier | en_US |
dc.rights | Copyright Elsevier | |
dc.source.uri | http://dx.doi.org/10.1016/j.jviromet.2004.09.008 | |
dc.subject | SARS-CoV | en_US |
dc.subject | Ectodomain | en_US |
dc.subject | CHO | en_US |
dc.subject | EGFP-tagged | en_US |
dc.subject | ACE2 | en_US |
dc.subject | Cell-based assay | en_US |
dc.title | A novel cell-based binding assay system reconstituting interaction between SARS-CoV S protein and its cellular receptor | en_US |
dc.type | Article | en_US |
dc.privacy.showsubmitter | false | |
dc.status.ispeerreviewed | true | |
dc.description.accreditation | Web of Science | en_US |