Fig. 7.
Biophysical and functional characterisation of B10v5 × hu225M with or without mutations in the CH1:CL interface. Analytical SEC after protein A purification of bsAbs produced in (A) HEK293-6E or (B) bsAbs produced in ExpiCHO after preparative SEC. The area under the curve in percent of all detected peak areas is given in the graph. (C) Simultaneous binding of both antigens cMET and EGFR using biolayer interferometry. BsAbs (wt, MaB40, MaB5/40, MaB21/40, MaB45/40) or antibodies consisting of only one Fab (oa for ‘one-armed’) were allowed to bind to cMET coated biosensors. Subsequently, biosensors were incubated with EGFR. (D) Differential scanning calorimetry of wildtype, interface mutants and ‘one-armed’ constructs (Table III). (E) Western blot analysis of the inhibition of EGFR and cMET phosphorylation in A549 cells by bsAbs with our without interface designs.