Virus-coated layer-by-layer colloids as a multiplex suspension array for the detection and quantification of virus-specific antibodies.

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 approximately 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.

journal_name

Clin Chem

journal_title

Clinical chemistry

authors

Toellner L,Fischlechner M,Ferko B,Grabherr RM,Donath E

doi

10.1373/clinchem.2005.065789

subject

Has Abstract

pub_date

2006-08-01 00:00:00

pages

1575-83

issue

8

eissn

0009-9147

issn

1530-8561

pii

clinchem.2005.065789

journal_volume

52

pub_type

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