The application of IgM in an xMAP suspension assay using recombinant Alt a1 as a model target protein
by Makonnen Abebe, Kathy Nguyen, Azam Tayabali, Wendy Decker, Veena Kumar, Serdal Sevinc, Hari Vijay
International Journal of Immunological Studies (IJIS), Vol. 1, No. 1, 2009

Abstract: The potential of Immunoglobulin M (IgM) in an Multi Analyte Profiling (xMAP) multiplex suspension assay was determined using Alt a1, a major allergen of Alternaria alternata, as the model target protein. LumAvidin (LAM) and carboxylated (CAM) microspheres, from LUMINEX™ Corporation, were compared. The reproducible standard curves for concentrations between 1 pg/mL and 10 ng/mL and 3 pg/mL and 300 ng/mL of the allergen were generated by the LAM assay, allowing for the quantitation of an unknown test sample. The success of the LAM assay is dependent on avidin molecules, initially bound on the microsphere, along with the use of a phycoerythrin-coupled secondary IgM. The standard curves generated by the CAM assay for the two allergen concentrations were erratic, falling short of quantifying the test sample. This is the first time a monoclonal IgM is successfully applied as a primary and secondary antibody in the xMAP format. The routine application of this assay will be consolidated as new IgM-based xMAP diagnostics become available.

Online publication date: Sat, 28-Feb-2009

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