Preparation and characterization of monoclonal antibodies against S1domain at N-terminal residues 249 to 667 of SARS-associated coron-avirus S1 protein

Abstract

Abstract: Objective To prepare and characterize monoclonal antibodies (mAbs) against S1 protein of severe acute respirato-ry 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 e xpressed in E.coli. The im-munogenicity of this S1 domain was identified and use d 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 im munosorbent assay (ELISA), indirect immunofluorescence assay (IFA) and Western blotting, respectively.Results Three hybridomas producing mAbs spe-cific 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 wer e IgG2a isotype, and the other was IgG1.Conclusion This is the first re-port of mAbs produced against S-protein receptor-binding domain of SARS-CoV. The 3 S1-s pecific mAbs may be useful for further study of the function of the S protein a nd for diagnosis of SARS-CoV infection.

WEN Kun¹, MEI Ya-bo¹, QIU Li-wen¹, LIAO Zhi-yong¹, Yuen Kwok-yung², CHE Xiao-yan¹. Preparation and characterization of monoclonal antibodies against S1domain at N-terminal residues 249 to 667 of SARS-associated coron-avirus S1 protein[J]. Journal of Southern Medical University, 2004, 24(01): 1-6.

References

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