Fig. 1
Characterization of the B12 antibody in scFv and IgG format. a Schematic domain structure of B12 in scFv format (left), corresponding to a molecular weight of 25 kDa, as shown in SDS-PAGE (right) in both reducing (R) and non-reducing (NR) conditions, and monomeric scFv molecules were separated from dimers by b size-exclusion chromatography (S75 column). c Specific binding of the antibody to biotinylated-ECD of IL-7Rα is shown by enriched ELISA signal as compared with control streptavidin. d Binding kinetics were performed by SPR technology on a BIAcore machine using CM5 microsensor chip. e Schematic domain structure of B12 reformatted into IgG (left), expressed in mammalian CHO cells with a molecular mass of 150 kDa (right) and purified by f size-exclusion chromatography (S200 column). g Improved ELISA signal toward IL-7Rα and h increased SPR signal as compared to the scFv c, d). i Representative snapshot from 0.5 μs MD simulations in explicit water. The amino acids that constitute the CDR of the antibody are in magenta. A movie of the MD simulations can be found in the online Supplementary Information
