title: Preparation and characterization of monoclonal antibodies against S1 domain at N-terminal residues 249 to 667 of SARS-associated coronavirus S1 protein.
abstract: OBJECTIVE
  To prepare and characterize monoclonal antibodies (mAbs) against S1 protein of severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV).
  
  
  METHODS
  6-His-tagged recombinant fragment at N-terminal residues 249 to 667 of SARS-CoV S1 protein including S-protein receptor-binding domain was expressed in E.coli. The immunogenicity of this S1 domain was identified and used to immunize BALB/c mice for the production of hybridomas. The identification of the mAbs against this S1 domain was performed using indirect enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA) and Western blotting, respectively.
  
  
  RESULTS
  Three hybridomas producing mAbs specific to the S1 domain was obtained, with a relative molecular mass of 48,500. None of the 3 mAbs were reactive with human coronaviruses 229E and OC43. Two of the mAbs were IgG2a isotype, and the other was IgG1.
  
  
  CONCLUSIONS
  This is the first report of mAbs produced against S-protein receptor-binding domain of SARS-CoV. The 3 S1-specific mAbs may be useful for further study of the function of the S protein and for diagnosis of SARS-CoV infection.
authors.affiliation: Central Laboratory, Zhujiang Hospital, First Military Medical University, Guangzhou 510282, China.
authors.forename: Kun
authors.initials: K
authors.lastname: Wen
authors.forename: Ya-bo
authors.initials: YB
authors.lastname: Mei
authors.forename: Li-wen
authors.initials: LW
authors.lastname: Qiu
authors.forename: Zhi-yong
authors.initials: ZY
authors.lastname: Liao
authors.forename: Kwok-yung
authors.initials: KY
authors.lastname: Yuen
authors.forename: Xiao-yan
authors.initials: XY
authors.lastname: Che
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D015153:Blotting, Western; D002460:Cell Line; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D008562:Membrane Glycoproteins / Q000276:immunology* / Q000502:physiology; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000276:immunology*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000737:chemistry*; D045169:Severe Acute Respiratory Syndrome / Q000175:diagnosis; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins / Q000276:immunology* / Q000502:physiology
chemical_list: D000911:Antibodies, Monoclonal; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins; C578557:spike glycoprotein, SARS-CoV; C578553:spike protein, mouse hepatitis virus