Wang, B. et al. Nat. Biotechnol. https://doi.org/10.1038/nbt.4052 (2018).

Next-generation sequencing has boosted the scale of immune profiling, by capturing the repertoire of T and B cell receptors that are enriched during adaptive immunity. In contrast, methods to profile antibody specificity typically require the laborious immortalization and in vitro expansion of individual B cells. Wang et al. are now accelerating the functional characterization of antibodies by using yeast engineered to display the human antigen-binding fragment (Fab) on its surface. To capture pairs of light and heavy chain antibody sequences from the same cell, they perform single B cell emulsion lysis followed by oligo-dT RNA capture, reverse transcription and overlap-extension PCR to create a single linked amplicon. The amplicon is then cloned into a yeast surface expression vector in a single step. The researchers used their method to identify neutralizing antibodies to HIV-1, Ebola virus glycoprotein and influenza hemagglutinin.