Using next-generation sequencing for discovery of high-frequency monoclonal antibodies in the variable gene repertoires from immunized mice.

Historically, isolation of antigen-specific monoclonal antibodies has relied on screening-based approaches. Here we describe a simple and rapid method for antibody isolation without screening, which capitalizes on next-generation DNA sequencing and bioinformatic analysis of antibody variable region (V) gene repertoires from the bone marrow plasma cells of immunized mice. The highest frequency antibody variable heavy (VH) and variable light (VL) gene sequences are paired based on their relative frequency, and their respective antibody genes are constructed by DNA synthesis and recombinantly expressed, purified, and validated for antigen-binding specificity.