title: Cleavage and serum reactivity of the severe acute respiratory syndrome coronavirus spike protein.
abstract: Severe acute respiratory syndrome (SARS) coronavirus (SCoV) spike (S) protein is the major surface antigen of the virus and is responsible for receptor binding and the generation of neutralizing antibody. To investigate SCoV S protein, full-length and individual domains of S protein were expressed on the surface of insect cells and were characterized for cleavability and reactivity with serum samples obtained from patients during the convalescent phase of SARS. S protein could be cleaved by exogenous trypsin but not by coexpressed furin, suggesting that the protein is not normally processed during infection. Reactivity was evident by both flow cytometry and Western blot assays, but the pattern of reactivity varied according to assay and sequence of the antigen. The antibody response to SCoV S protein involves antibodies to both linear and conformational epitopes, with linear epitopes associated with the carboxyl domain and conformational epitopes associated with the amino terminal domain. Recombinant SCoV S protein appears to be a suitable antigen for the development of an efficient and sensitive diagnostic test for SARS, but our data suggest that assay format and choice of S antigen are important considerations.
authors.affiliation: School of Animal and Microbial Sciences, University of Reading, Reading, United Kingdom.
authors.forename: Yong Xiu
authors.initials: YX
authors.lastname: Yao
authors.forename: Junyuan
authors.initials: J
authors.lastname: Ren
authors.forename: Paul
authors.initials: P
authors.lastname: Heinen
authors.forename: Maria
authors.initials: M
authors.lastname: Zambon
authors.forename: Ian M
authors.initials: IM
authors.lastname: Jones
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000276:immunology; D015153:Blotting, Western; D002478:Cells, Cultured; D005434:Flow Cytometry; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D045473:Severe acute respiratory syndrome-related coronavirus / Q000276:immunology* / Q000378:metabolism; D045169:Severe Acute Respiratory Syndrome / Q000175:diagnosis / Q000276:immunology; D064370:Spike Glycoprotein, Coronavirus; D018411:Spodoptera; D014357:Trypsin / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins; C578557:spike glycoprotein, SARS-CoV; C578553:spike protein, mouse hepatitis virus; D014357:Trypsin