Received on December 22, 2005
Accepted on May 8, 2006

Automation and Analytical Techniques

Virus-Coated LbL Colloids as a Multiplex Suspension Array for the Detection and Quantification of Virus-Specific Antibodies

¹ Institute of Medical Physics and Biophysics, Leipzig University, Leipzig, Germany
² Institute of Applied Microbiology, University of Natural Resources and Applied Life Sciences, Vienna, Austria

^* To whom correspondence should be addressed. E-mail: done{at}medizin.uni-leipzig.de.

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 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.