ON-AIRRseq: open-sourced nanopore-based antibody immune receptor repertoire sequencing 4212

Abstract Description Monoclonal antibodies (mAbs) are indispensable in diagnostics, therapeutics, and research. However, traditional methods for studying antibody immune receptor repertoires (AIRR) in response to disease or immunization are often prohibitively expensive, time-intensive, and reliant on specialized equipment. These barriers have limited global access to AIRR analysis, hindering advancements in education and research. To overcome these challenges, we developed ON-AIRRseq, an open-source Nanopore-based AIRR sequencing platform. This innovative system uses customizable forward primers with barcodes and unique molecular identifiers, enabling gene expression comparisons across multiple samples in a single run. A transcript-specific reverse primer ensures rapid adaptability between isotypes or other desired targets. Validation of ON-AIRRseq in six murine hybridomas demonstrated its ability to recover over 50 reads per sequence, identify aberrant IGHV and IGKV genes, and outperform commercially available kits in both efficiency and adaptability. Current efforts are focused on expanding the system’s scalability to analyze antibody responses in murine vaccination studies. ON-AIRRseq offers a cost-effective, highly adaptable, and scalable approach to repertoire sequencing, designed to improve accessibility for education and research worldwide. Funding Sources This project was supported by grants from the National Institutes of Health (NIH), National Institute of General Medical Sciences (NIGMS), IDeA Networks of Biomedical Research Excellence (INBRE), Award number: P20GM103466. The content is solely the authors’ responsibility and does not necessarily represent the official views of the National Institutes of Health. Topic Categories Computational and Systems Immunology (COMP)