Objective

Interference by endogenous antibodies (heterophiles, HAMA, HAAA, RF, and others) in immunoassays is a well-documented phenomenon.  Remarkably, this still continues to plague the immunoassay industry. Multiplex assays such as Luminex® bead-based assays, can be particularly vulnerable to these effects and can leave the user with confusing, inaccurate data at best, and erroneous interpretations at worst. Ultimately, it is the responsibility of the manufacturer to produce an assay design that can manage these and other sample matrix effects to produce accurate data.  In this study, we demonstrate the profound effects that unresolved HAMA and RF can have on the data. We chose a standard Luminex assay from a leading supplier and tested for interferences from samples with known HAMA or RF levels and compared that performance to the R&D Systems® Human XL Cytokine Discovery Luminex Assay.  These results clearly demonstrate that development expertise and assay design can greatly impact the quality and accuracy of the data from multiplex assays by eliminating or greatly reducing effects from endogenous antibodies in the sample. xMAP® technology provides a convenient platform for assessing common immunoassay interferences by simultaneous evaluation, with the benefits of speed and efficiency.