Figure 1
Confirmation of antibody integrity and antigen binding in SPR. (A) HPLC-SEC chromatograms of the IgG variants as relative UV280 nm signal normalized to maximum response. The chromatograms of molecules overlap. (B) SDS-PAGEs under non-reducing (NR) and reducing (R) conditions. Shown are two independently prepared SDS-PAGEs of purified V-gene matched IgG variants (left) and the IgE supernatant (right). The uncropped original images showing the relevant gel parts can be found in Supplementary Fig. 1A and B, respectively. (C) IgE-specific Western Blot after native PAGE of anti-biotin IgE supernatant next to contact allergy serum sample32. The uncropped original image showing the relevant lanes can be found in Supplementary Fig. 1C. (D) Schematic overview of SPR experiments showing anti-biotin antigen binding and presence of human IgE in culture supernatants. (E) Sensorgrams of one representative of three independent experiments showing binding of anti-IgE antibody (left) and soluble FcεRI (right) at pH 7.4 and pH 6.0 after capturing anti-biotin IgE from culture supernatant in comparison to IgG at 100 nM.
