DNA sequencing technologies, especially NGS, are extensively applied in the process of antibody discovery and development. Techniques like hybridoma sequencing and phage and yeast display technology that rely on NGS or nucleotide sequencing methods have strengthened the development of antibody therapeutics and diagnostics.
Hybridoma sequencing, a type of nucleotide sequencing, can obtain sequence information of monoclonal antibodies produced by hybridoma cell lines, widely used for the production of humanized antibodies, and in the initial screening and lead selection. Display technologies rely on NGS to generate libraries of antibody sequences that are then expressed or displayed in large numbers on the surface of phages, yeast, or bacteria, allowing the rapid identification of antibodies and fragments that bind target molecules.
NGS may also be used for recombinant expression of antibody clones through two means: single B cell sequencing or B cell repertoire sequencing (BCR), acting as a powerful tool to analyze large-scale sequence data of antibody repertoires in response to particular antigens or diseases directly from blood B cells.
Single B cell sequencing relies on the individual culturing of B cells prior to sequencing, retaining information on light and heavy chain pairing. BCR sequencing depends on the bulk mRNA extraction of B cells, particularly from the spleen. Both help researchers gain important knowledge of the humoral adaptive immune response to identify therapeutics and monitor disease in clinical settings.