(115) Dual Reporter multiplexing enables simultaneous detection of IgG and IgM antibodies against autoantigens and pathogenic proteins in human samples

Introduction/Rationale: Profiling both IgG and IgM antibodies against autoantigens and infectious disease targets provide insight into the immune response and disease progression. IgM antibodies are typically associated with an early immune response, while IgG antibodies are suggestive of more chronic or persistent immune processes.

Methods: Using the xMAP® INTELLIFLEX® DR-SE system, which features a second reporter channel, we developed a Dual Reporter IgG/IgM Reagent Pack that can be used with any of the MILLIPLEX® serology kits to measure the immune response to various disease-relevant targets, specifically detecting IgG and IgM antibodies on separate channels for each multiplexed antigen bead. The Dual Reporter Reagent Pack was used to profile 88 antigens across 5 existing serology-based MILLIPLEX® kits in apparently healthy and disease serum and plasma samples. Further, to demonstrate the relevance of the Dual Reporter Reagent Pack for vector-borne disease studies, we compared serum and plasma from apparently healthy donors (n=32) versus individuals with Lyme Disease (n=8) using a novel multiplex panel containing the Borrelia antigens OspA, OspB, OspC, Flagellin, and VlsE1.

Results: Our results demonstrated significantly increased binding of both IgG and IgM antibodies to OspC and VlsE1 proteins in Lyme Disease samples. Additionally, IgG binding to the OspB protein was also elevated in Lyme Disease compared to healthy control samples.

Conclusion: In summary, the Dual Reporter IgG/IgM Reagent Pack enhances the capability for simultaneous profiling of IgG and IgM antibodies, which is applicable to multiple disease states including cancer, neurodegeneration, autoimmune disease, and Lyme Disease.