(440a) High Throughput Sequencing of Antibody Heavy and Light Chain Pairings

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. 
V_H-V_L 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.