Fig. 1: Overview of the evaluation system for epitope similarity and the Epitope Binning-seq platform. | Communications Biology

Fig. 1: Overview of the evaluation system for epitope similarity and the Epitope Binning-seq platform.

From: Epitope binning for multiple antibodies simultaneously using mammalian cell display and DNA sequencing

Fig. 1

a Cell surface display of the query antibody (qAb). The cDNA encoding a single chain variable fragment (scFv) of qAb is transduced into the antigen-expressing cells. b Evaluation of qAbs sharing a similar epitope with the fluorescently labeled reference antibody (rAb). Because the target epitope on the antigen is masked by qAb, the rAb cannot bind to the antigen and an rAb-non-binding [rAb(−)] cell population is detected in FCM analysis. Forward scatter, FS. c Evaluation of qAbs with a distinct epitope to that of rAb. The rAb binds to the target epitope and FCM analysis reveals an rAb-binding [rAb(+)] cell population. d Parallel antibody classification via Epitope Binning-seq. An antigen-expressing qAb cell library is incubated with rAbs, which bind to distinct epitopes, and each rAb(−) cell population is sorted for NGS analysis to reveal qAb sequences. By analyzing qAbs enriched in the corresponding rAb(−) populations, the qAbs can be classified into different epitope bins, which are composed of qAbs with epitopes similar to each rAb.

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