2.6 BLI-based binding IgG immune-complexes
A comparison of binding kinetics between IgG or IgG in complex with
dimeric Fel d 1 and FcγRIIb (CD32b) was also performed using BLI assays.
Briefly, SAX biosensors were saturated with 2 µg/ml of human recombinant
biotinylated FcγRIIb (CD32b) followed by association with 25 nM of
either monomeric IgG1 and IgG4 F127 or in pre-complexed form with 25 nM
Fel d 1 for 300 s. Dissociation was also performed in 300 s in kinetics
buffer. Drift control was performed with a loaded sensor dipped in
kinetics buffer only. The resulting data were normalized to the highest
response, using OriginPro (OriginLab Corporation, Northampton, MA, USA)
software analysis