Rapid discovery of high-affinity antibodies via massively parallel sequencing, ribosome display and affinity screening.

ABSTRACT: Developing therapeutic antibodies is laborious and costly. Here we report a method for antibody discovery that leverages the Illumina HiSeq platform to, within 3 days, screen in the order of 10⁸ antibody-antigen interactions. The method, which we named 'deep screening', involves the clustering and sequencing of antibody libraries, the conversion of the DNA clusters into complementary RNA clusters covalently linked to the instrument's flow-cell surface on the same location, the in situ translation of the clusters into antibodies tethered via ribosome display, and their screening via fluorescently labelled antigens. By using deep screening, we discovered low-nanomolar nanobodies to a model antigen using 4 × 10⁶ unique variants from yeast-display-enriched libraries, and

SUBMITTER: Porebski BT 

PROVIDER: S-EPMC10963267 | biostudies-literature | 2024 Mar

REPOSITORIES: biostudies-literature