Fluorescence-activated droplet sequencing (FAD-seq) directly provides sequences of screening hits in antibody discovery.

ABSTRACT: Droplet microfluidics has become a very powerful tool in high-throughput screening, including antibody discovery. Screens are usually carried out by physically sorting droplets hosting cells of the desired phenotype, breaking them, recovering the encapsulated cells, and sequencing the paired antibody light and heavy chain genes at the single-cell level. This series of multiple consecutive manipulation steps of rare screening hits is complex and challenging, resulting in a significant loss of clones with the desired phenotype or large fractions of cells with incomplete antibody information. Here, we present fluorescence-activated droplet sequencing, in which droplets showing the desired phenotype are selectively picoinjected with reagents for RT-PCR. Subsequently, light and heavy chain gene

SUBMITTER: Autour A 

PROVIDER: S-EPMC11406258 | biostudies-literature | 2024 Sep

REPOSITORIES: biostudies-literature