Fig. 2

Structural stability of the recombinant antibodies. Stability of RmAb158, Tetra-RmAb158 and Hexa-RmAb158 under thermal stress evaluated with Tycho nt.6 system (NanoTemper Technologies, München, Germany). Inflection temperature(s) at which the proteins destabilize measured for the three antibodies (final concentration of 1 μM) at a 0 h, b 2 h, and c 72 h incubation at 37 °C. Results presented as the ratio of fluorescence intensity at 350 nm to 330 nm. Changes in fluorescence intensities reflect changes in the proteins’ structural stability. Due to the presence of scFv on their N-terminal ends, both Tetra-RmAb158 and Hexa-RmAb158 displayed lower inflection temperatures compared to RmAb158. Inflection temperatures of the three antibodies remained constant after 2 h and 72 h incubation at 37 °C, further suggesting structural stability of these antibodies. Results are generated from two-repetitive experiments and average values are shown