Multiplex immuno screening assay
AssigneeInstitut Pasteur
InventorsJean-Claude Manuguerra, Jessica Vanhomwegen, Philippe Despres, Sylvie Paulous
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Abstract
The present invention provides an immunoassay leading to the rapid and simultaneous detection of antibodies to a wide range of infectious pathogens in biological fluids of infected patients. This immunoassay involves the covalent and oriented coupling of fusion proteins comprising an AGT enzyme and a viral antigen on an identifiable solid support (e.g. fluorescent microspheres). The thus obtained antigen-coupled microspheres show enhanced capture of specific antibodies as compared to antigen-coupled microspheres produced by standard amine coupling procedures.
Claims9
An in vitro assay method for detecting at least two different target antibodies present in a biological sample from a subject, said method comprising the steps of:
- (a) contacting a mixture of at least 2 solid supports with a biological sample from a subject comprising a mixture of human antibodies;
- wherein one of the solid supports comprises a first 6-alkylguanine-DNA-alkyltransferase-antigen fusion protein covalently coupled to a 6-alkylguanine-DNA-alkyltransferase-antigen substrate, and
- wherein another of the solid supports comprises a second 6-alkylguanine-DNA-alkyltransferase-antigen fusion protein covalently coupled to a 6-alkylguanine-DNA-alkyltransferase-antigen substrate;
- wherein said antigen is an epitope of a viral protein from a virus selected from:
- Dengue virus 1, Dengue virus 2, Dengue virus 3, Dengue virus 4, West Nile virus, Yellow Fever virus, Japanese encephalitis virus of genotype 1, 2, 4, or 5, Zika virus, Wesselbron virus, Rocio virus, Murray encephalitis virus Saint-Louis encephalitis virus, and Rabensburg virus;
- (b) detecting the presence or absence of binding of antibodies to the first 6-alkylguanine-DNA-alkyltransferase-antigen fusion protein by detecting the binding or lack of binding of one of the two target antibodies to the first fusion protein;
- and
- (c) detecting the presence or absence of binding of antibodies to the second 6-alkylguanine-DNA-alkyltransferase-antigen fusion protein by detecting the binding or lack of binding of the second of the two target antibodies to the second fusion protein.
Dependencies are inferred from the claim text.