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David Onley

Publications and source records attributed to David Onley.

2 recordsLinked to original sources

A comparability study of the emerging protein array platforms with established ELISA procedures.

Recent developments in microarray technology have made it possible to perform immunoassays in a multiplexed format. This ability is highly desirable given the potential for greater throughput analysis. In spite of the obvious advantages, a number of issues arise as a result of multiplexing the reactions. In this study, we have assessed the performance characteristics that are associated with the transfer of technology from a uniplexed to a multiplexed format. Two solution phase array platforms were chosen for this study: the commercially available Luminex(100) xMap system (Austin, Texas, USA) and the UltraPlex technology devised by SmartBead Technologies Ltd. (Cambridge, UK). For this comparative study, a test for the presence of six autoantibodies in a selection of human patient serum samples was chosen as a model system. The multiplexed Luminex xMap and SmartBead UltraPlex assays were generally comparable. However, both systems generated some results that were at variance with those obtained by ELISA. The different methods used for the assignment of the cut-off levels for each of the assays within any given platform was identified as the major source of these non-concordant results. The present study demonstrates that array platforms have the potential to be used in immunodiagnostics providing that suitable cut-off levels are established.

Antibodies↗

Determination of ANA specificity using the UltraPlex platform.

SmartBead Technologies has developed a multiplexed immunofluorescence assay, the UltraPlex ANA Profile, which determines nine antinuclear antibodies simultaneously. The UltraPlex assay platform uses bar-coded microparticles to track analytes through assays. These bar-coded microparticles were used to tag and track key ANA markers: Jo-1, Scl-70, Sm, SmRNP, SSA, SSB, U1RNP, Centromere B, dsDNA, and a blank control microparticle. The immunofluorescence assays are fully automated and are performed on a Perkin-Elmer multiprobe II liquid handling system that performs all sera dilutions, additions of reagents, washes, and incubation steps. Results were determined by the automated UltraPlex plate reader. This fully automated multiplex antinuclear antibody (ANA) immunoassay was used to screen commercially available ANA-positive sera and negative control samples. The UltraPlex ANA Profile enables the panels of samples to be screened simultaneously for nine ANA antoantibodies, requiring significantly less labor and fewer reagents, with performance equivalent to existing gold-standard methods.

Antibodies, Antinuclear↗