( LaRene Kuller David An...)Development of a Multiplex Bead-Based Assay for Antibody Screening of a Nonhuman Primate Colony on the Bio-Plex System

LaRene Kuller, David Anderson, Dominic Andrada, and Richard Grant, National Primate Research Center, University of Washington, Seattle, WA 98195 USA

Introduction
We have been using four separate enzyme-linked immunosorbent assays (ELISAs) to screen our primate colony for the viruses of greatest concern: simian retrovirus (SRV), simian T-cell leukemia virus (STLV), herpes B, and simian immunodeficiency virus (SIV). To minimize the labor involved, testing of antibody responses to these viruses requires that these individual assays be performed on a limited sample volume. As we add more viruses to our screening protocol, serological testing by ELISA will become more time-consuming.

The Bio-Plex suspension array system, which is based on Luminex xMAP technology, is a multiplex flow cytometricbased system that utilizes up to 100 color-coded 5.6 m polystyrene bead sets. Each bead set is internally dyed with different ratios of two spectrally distinct fluorophores. Each of the bead sets can be conjugated with a unique protein, peptide, antibody, or, in our case, disrupted whole virus. The conjugated beads are pooled together in the wells of a microplate with the sample to be tested (plasma), followed by the addition of a detection antibody, forming a capture immunoassay that is then read by the Bio-Plex suspension array reader. Each separate reaction is identified and quantitated based on the bead color. This allows a single sample to be tested simultaneously for reaction to a number of different viruses.

Our goal was to determine whether the Bio-Plex system yields results comparable to those of our current ELISA assays. The capability to multiplex would allow us to
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