Recombinant Human B Cell Repertoires Enable Screening for Rare, Specific, and Natively Paired Antibodies

Kierny MR, et al., doi:10.1038/s42003-017-0006-2, Nature Communications Biology, 2018

Described herein is the encapsulation of primary B cells into water-in-oil droplets, where their cognate VH and VL domains were amplified and paired to form scFv libraries. Furthermore, this approach was used to generate natively paired phage-display libraries from healthy individuals and to enrich for cross-reactive antibodies targeting influenza hemagglutinin (HA). Bio-Layer Interferometry (BLI) was used to study the binding affinities and to perform an epitope binning assay. ScFv-Fc antibodies were immobilized onto Protein A (ProA) Biosensor probes. Antibody loaded ProA sensor tips were dipped in 5 concentrations of biotinylated hemagglutinin H1 or hemagglutinin H5 for an association step, followed by a dissociation step in buffer. Binding affinities of certain antibodies were studied using Anti-human Fc biosensors. Data obtained were fitted using a 1:1 binding model. Equilibrium dissociation constants (KD) were determined. Competition assays were performed by dipping biotinylated antibody loaded Streptavidin (SA) Biosensor tips in wells containing hemagglutinin H1, followed by incubation with a competing IgG. Collectively, the method developed in this study can be used to rapidly screen for rare, specific, and natively paired antibodies from millions of primary human B cells.

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