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. 2016 Apr 22;473(1):23-28.
doi: 10.1016/j.bbrc.2016.03.038. Epub 2016 Mar 12.

Identification of Antigen-Specific Human Monoclonal Antibodies Using High-Throughput Sequencing of the Antibody Repertoire

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Identification of Antigen-Specific Human Monoclonal Antibodies Using High-Throughput Sequencing of the Antibody Repertoire

Ju Liu et al. Biochem Biophys Res Commun. .

Abstract

High-throughput sequencing of the antibody repertoire provides a large number of antibody variable region sequences that can be used to generate human monoclonal antibodies. However, current screening methods for identifying antigen-specific antibodies are inefficient. In the present study, we developed an antibody clone screening strategy based on clone dynamics and relative frequency, and used it to identify antigen-specific human monoclonal antibodies. Enzyme-linked immunosorbent assay showed that at least 52% of putative positive immunoglobulin heavy chains composed antigen-specific antibodies. Combining information on dynamics and relative frequency improved identification of positive clones and elimination of negative clones. and increase the credibility of putative positive clones. Therefore the screening strategy could simplify the subsequent experimental screening and may facilitate the generation of antigen-specific antibodies.

Keywords: Antibody repertoire; Antigen-specific human monoclonal antibodies; High-throughput sequencing; Screening strategy.

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