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    Yann Chong Tan.
    Despite tremendous progress in understanding immune responses, the ability to characterize functional antibody responses remains limited. I developed a method that utilizes barcoding to enable high-throughput sequencing of paired heavy-chain and light-chain immunoglobulin sequences. This approach enables us to generate dendrograms of functional affinity-matured antibody responses, and express selected antibodies for functional characterization. This method was applied to a human who received an influenza vaccine, humans with Staphylocuccus aureus infection, a human with rhuematoid arthritis, an autoimmune disease, and a human who has lung adenocarcinoma, and is a long term non-progressor. These are examples in vaccination, infection, autoimmune disease and cancer, respectively. In each case, we show that we were able to obtain paired heavy and light chain sequences. We expressed selected antibodies for functional characterization and showed that we were able to obtain functional antibodies that bound to target antigens in each of the above indications. Furthermore, we also showed that large clonal families dominate in influenza vaccination, and that larger antibody clonal families have higher binding affinities. Taken together, the approach provides a method to directly characterize functional antibody responses that could advance our understanding of protective and pathogenic immune responses in health and disease.
    Digital Access   2012