273449 Award Submission: High Throughput Sequencing of Antibody Heavy and Light Chain Pairings

Tuesday, October 30, 2012: 5:21 PM
407 (Convention Center )
Brandon DeKosky1, Ryan P. Deschner1, Brandon Rawlings1, Gregory Ippolito1, C. Grant Willson2, Andrew D. Ellington3 and George Georgiou4, (1)Department of Chemical Engineering, The University of Texas at Austin, Austin, TX, (2)McKetta Department of Chemical Engineering, The University of Texas at Austin, Austin, TX, (3)Department of Biochemistry, University of Texas at Austin, Austin, TX, (4)Departments of Chemical Engineering and Biomedical Engineering, The University of Texas at Austin, Austin, TX

Next generation DNA sequencing (NGS) is transforming the field of immunology by giving researchers the potential to sequence immune repertoires at unprecedented depth.  However, the utility of NGS for repertoire analysis is severely curtailed by its inability to resolve the pairing between antibody heavy and light chains and T-cell receptor α and β strands.  To directly address this issue, we have developed an emulsion-based method to sequence natively paired antibody heavy and light chains at high-throughput.  VH-VL pairings have been validated using mixtures of various immortalized B-cell clones with known antibody sequences to confirm accuracy.  High-throughput sequencing of the heavy and light chain pairing is a significant step toward greater understanding of the immune repertoire, and potential applications include immune response analysis and antibody discovery.

This work was supported by the University of Texas at Austin Cockrell School of Engineering, the Harrington Foundation, the National Science Foundation, and The Hertz Foundation.


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