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Toellner, L; Fischlechner, M; Ferko, B; Grabherr, RM; Donath, E.
(2006): Virus-coated layer-by-layer colloids as a multiplex suspension array for the detection and quantification of virus-specific antibodies.
Clin Chem. 2006; 52(8):1575-1583 FullText FullText_BOKU

Abstract:: Background: Suspension array technology has surpassed ELISA for automated, simultaneous detection and quantification of soluble biomarkers such as virus-specific antibodies. We describe assays in which antigens are attached to a lipid bilayer surrounding color-coded particles. Methods: We used layer-by-layer technology to establish a multiplex suspension array with distinguishable microbeads coated with authentic viral surfaces to catch and quantify virus-specific antibodies in a flow cytometric analysis. Antigenic surfaces were generated by chimeric and wild-type baculoviruses plus 2 different influenza A virus subtypes fused to a lipid bilayer surrounding distinctly colored particles. Specificity of binding of chosen antibodies and sera was detected by immunofluorescence. Results of multiplex analysis were compared with results of ELISA. Results: Titrations of virus-specific antibodies in the multiplex suspension array demonstrated specific binding to the viral surface proteins. The multiplex suspension array gave positive results for up to log 5-diluted primary antibodies with an similar to 5- to 10-fold reduced dynamic range compared with the respective ELISA. Conclusions: The bead-based multiplex suspension array is customizable and easy to establish. By displaying native influenza A virus surfaces and recombinant HIV-1 epitopes, the new assay provides a tool for the detection of major viral infections in humans. (c) 2006 American Association for Clinical Chemistry.
Authors BOKU Wien:: Grabherr Reingard
Find related publications in this database (using NML MeSH Indexing): Antibodies, Viral - blood; Antigens, Viral - chemistry; Baculoviridae - chemistry; Colloids -; Enzyme-Linked Immunosorbent Assay -; Flow Cytometry -; Fluorescent Antibody Technique -; Hemagglutinin Glycoproteins, Influenza Virus - analysis; Humans -; Immunoassay - methods; Influenza A virus - chemistry; Lipid Bilayers - chemistry; Microspheres -; Viral Fusion Proteins - analysis