Fig. S2.
Kinetic evaluation of 4LB5 binding to recombinant NCL and 4LB5 specific binding to NCL. (A) 4LB5 affinity for recombinant NCL-RBD was assessed by SPR using increasing concentrations (1–50 nM) of scFv. Resulting Kd is also indicated. (B) The detection limit of ELISA using 4LB5 (see Fig. 1) was assessed using different amounts of scFv and indicated numbers of MDA-MB-231 cells. Data (normalized for background levels) are representative of two independent experiments performed in triplicate, ± SD, **P < 0.01, compared with the corresponding negative control. (C) Specific binding of 4LB5 to NCL assessed by Western Blot on MDA-MB-231 cells transfected with control (siCTRL) or anti-NCL (siNCL) siRNA. 4LB5 was used as a primary antibody and HRP-conjugated anti-His6 as secondary antibody. Bands indicated by the asterisk are caused by the secondary anti-His6 antibody used for the detection of the NCL-bound scFv. Tubulin was used as a loading control.