Abstract
Biomarker quantitation by Ligand Binding Assays is susceptible to interference. Such interference usually manifests as a signal decrease resulting in target underestimation. However, a recently developed biomarker assay for soluble CD137 exhibited an uncharacteristic positive interference in presence of the therapeutic antibody urelumab (anti-CD137). Urelumab was found to be capable of formation of higher order complex through simultaneous multivalent interactions with both soluble CD137 and assay reagents. Higher Order Complex formation by urelumab was qualitatively confirmed by light scattering methods. This Higher Order Complex network enhanced the signal output leading to overestimation of the soluble CD137 analyte. Thus the positive interference in the soluble CD137 biomarker assay appears to be caused by the therapeutic antibody-mediated formation of a Higher Order Complex network.
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