Figure 1.

Workflow summary of mAb discovery pipeline in mice. Upon completion of the immunization regimen, spleens are harvested and gently homogenized to a single-cell suspension. The cell suspension is treated by negative magnetically-assisted selection until predominantly B cells remain. The B-cell suspension is stained using a cocktail of antibodies and Ag-streptavidin (as well as decoy-streptavidin) complexes to facilitate the sorting of Ag-positive class-switched cells. The cells are then cultured and induced to express antibodies in presence of supportive cytokines and feeder cells. Culture-conditioned supernatants containing target antibodies are then screened using Ag-specific ELISA, and positive wells are harvested for RNA isolation. Ab-encoding RNA is then reverse-transcribed, amplified and cloned into an expression cassette enabling recombinant production of the mAbs, followed by functional validation.