Decoding the Antibody Reactome at Scale with Novel Antigen Display Technology to Decipher Immune History 2335216

Abstract Introduction The human immune system serves as a ledger of environmental exposures, infections, and self-reactivity and there is a there is a critical need for high-throughput tools capable of mapping the entire human antibody reactome (all the antibody-antigen interactions). However, capturing the entire “immune history” can be difficult as conventional assays are limited to a narrow set of defined antigens. Methods Molecular Indexing of Proteins by Self-Assembly (MIPSA) technology employs a single-pot process to create libraries with antigens are covalently linked to unique DNA barcodes and decoded via Next-Generation Sequencing. This multiplexed, sample-sparing approach enables the simultaneous profiling of tens of thousands of interactions via core libraries: (1) HuSIGHT, covering the human proteome with 353,034 peptides and 16,516 full-length proteins; (2) VirSIGHT, covering all viruses known to infect humans with 285,669 peptides; and (3) EnviroSIGHT, covering 1,207 genera of allergens, microbial antigens, and toxins with 242,884 peptides. Results Using commercial monoclonal antibodies and serum from patients with confirmed autoimmune diseases, the inclusion of both peptides and full-length proteins increases the detection of autoantibody targets on HuSIGHT. Profiling with VirSIGHT effectively distinguished “common” viral signatures from unique exposure histories. Furthermore, EnviroSIGHT allowed for the discrimination of distinct allergic profiles and geographical locations based on endemic pathogen exposure of the participants. We validated the platform’s robustness against technical and clinical confounders. Conclusion The MIPSA technology offers a scalable, high-resolution solution for uncovering immune insights offering a comprehensive view of immune history, this platform facilitates the discovery of novel biomarkers, infectious triggers of chronic conditions, the characterization of host-pathogen interactions, and the development of more effective therapeutic interventions for human health. Funding Source n/a Topic Categories Technological Innovations in Immunology (TECH)