Fig. 7: Potent pneumovirus neutralizing antibodies identified from a human antibody library.
a Schematic representation of the phage display workflow. b Gene family analysis of eight scFv clones identified from a phage-displayed human antibody (scFv) library using RSV-F UFCR1-P2-NQ as a biopanning antigen. c ELISA-derived EC50 (µg/ml) values of library-derived antibodies binding to RSV-F sc9-10 DS-Cav1 and hMPV-F UFCM1-P2-iSS. d IC50 (µg/ml) values derived from live RSV and hMPV neutralization assays. Error bars in (c, d) represent the difference between duplicates at each concentration tested for each sample. e Distribution of germline gene usage, somatic hypermutation, and CDR3 length plotted for heavy chains (HCs) and λ-light chains (λ-LCs) of the scFv library during the biopanning process. f Identity-divergence analysis of the IXL-A4 (or A4) within the scFv library during the biopanning process. The sequence datasets used in (e) and (f) were obtained from next-generation sequencing (NGS) of the scFv libraries on an Ion GeneStudio S5 platform. For the heatmaps in (f), after data processing using an Antibodyomics pipeline, each sequence is plotted as a function of sequence identity from a reference antibody chain and sequence divergence from the assigned germline gene. Color indicates sequence density at a particular point on the 2D plot. Sequences with the same germline gene, a CDR3 identity ≥95%, and a 1-residue error margin of CDR length calculation relative to the reference antibody chain are plotted as orange dots with the number of sequences and gene family percentage labeled. The schematic representation of phage-based antibody isolation was created in BioRender. Lee, Y. (2024) BioRender.com/p25x130.
