Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 187, Issue 1, Pages 72-78Publisher
ELSEVIER
DOI: 10.1016/j.jviromet.2012.09.006
Keywords
SARS-CoV; Spike protein; Bi-specific monoclonal antibodies; Quadroma; ELISA
Funding
- National Institute of Allergy and Infectious Diseases, National Institutes of Health, USA [5U01AI061233 05]
- NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES [U01AI061233] Funding Source: NIH RePORTER
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The severe acute respiratory syndrome coronavirus (SARS-CoV) spike protein is known to mediate receptor interaction and immune recognition and thus it is considered as a major target for vaccine design. The spike protein plays an important role in virus entry, virus receptor interactions, and virus tropism. Sensitive diagnosis of SARS is essential for the control of the disease in humans. Recombinant SARS-CoV S1 antigen was produced and purified for the development of monoclonal and bi-specific monoclonal antibodies. The hybridomas secreting anti-S1 antibodies, F26G18 and P136.8D12, were fused respectively with the YP4 hybridoma to generate quadromas. The sandwich ELISA was formed by using F26G18 as a coating antibody and biotinylated F26G18 as a detection antibody with a detection limit of 0.037 mu g/ml (p < 0.02). The same detection limit was found with P136.8D12 as a coating antibody and biotinylated F26G18 as a detection antibody. The sensitivity was improved (detection limit of 0.019 mu g/ml), however, when using bi-specific monoclonal antibody (F157) as the detection antibody. In conclusion, the method described in this study allows sensitive detection of a recombinant SARS spike protein by sandwich ELISA with bi-specific monoclonal antibody and could be used for the diagnosis of patients suspected with SARS. (C) 2012 Elsevier B.V. All rights reserved.
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