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Development and characterisation of neutralising monoclonal antibody to the SARS-coronavirus
- Source :
- Journal of Virological Methods
- Publication Year :
- 2004
- Publisher :
- Elsevier B.V., 2004.
-
Abstract
- There is a global need to elucidate protective antigens expressed by the SARS-coronavirus (SARS-CoV). Monoclonal antibody reagents that recognise specific antigens on SARS-CoV are needed urgently. In this report, the development and immunochemical characterisation of a panel of murine monoclonal antibodies (mAbs) against the SARS-CoV is presented, based upon their specificity, binding requirements, and biological activity. Initial screening by ELISA, using highly purified virus as the coating antigen, resulted in the selection of 103 mAbs to the SARS virus. Subsequent screening steps reduced this panel to seventeen IgG mAbs. A single mAb, F26G15, is specific for the nucleoprotein as seen in Western immunoblot while five other mAbs react with the Spike protein. Two of these Spike-specific mAbs demonstrate the ability to neutralise SARS-CoV in vitro while another four Western immunoblot-negative mAbs also neutralise the virus. The utility of these mAbs for diagnostic development is demonstrated. Antibody from convalescent SARS patients, but not normal human serum, is also shown to specifically compete off binding of mAbs to whole SARS-CoV. These studies highlight the importance of using standardised assays and reagents. These mAbs will be useful for the development of diagnostic tests, studies of SARS-CoV pathogenesis and vaccine development.
- Subjects :
- Monoclonal antibody
medicine.drug_class
Protein Conformation
viruses
Blotting, Western
Enzyme-Linked Immunosorbent Assay
medicine.disease_cause
Antibodies, Viral
Virus
Epitope
Article
Epitopes
Mice
SARS-CoV, human severe acute respiratory syndrome coronavirus
Antigen
Viral Envelope Proteins
Nidovirales
Neutralization Tests
Virology
Chlorocebus aethiops
medicine
Animals
mAb, monoclonal antibody
skin and connective tissue diseases
Antigens, Viral
Vero Cells
Coronavirus
Membrane Glycoproteins
biology
SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis
Immunochemistry
fungi
Antibodies, Monoclonal
biology.organism_classification
Nucleoprotein
body regions
Neutralising
Nucleoproteins
SARS-coronavirus
Severe acute respiratory syndrome-related coronavirus
Immunoglobulin G
Spike Glycoprotein, Coronavirus
biology.protein
Antibody
Subjects
Details
- Language :
- English
- ISSN :
- 18790984 and 01660934
- Volume :
- 120
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Journal of Virological Methods
- Accession number :
- edsair.doi.dedup.....0d15d3f951351f1c442ff2db87ef8d5e