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. 2016 Feb 5;291(14):7558–7570. doi: 10.1074/jbc.M115.701656

FIGURE 3.

FIGURE 3.

BLI assay for binding affinity measurements. Two highest affinity compounds, KMS31 and KMS32 (biotin tagged), were immobilized on streptavidin sensors. The kinetic measurements were carried out by exposing sensors with serially diluted soluble CLL 014 IgG in binding buffer (1× PBS, pH 7.4 containing 1% BSA) in wells of a 96-well microtiter plate. The association and dissociation profiles of the compounds was monitored at 30 °C using an Octet RED96 system (Pall ForteBio). Binding curves were analyzed by global fitting of sensorgrams to a 1:1 binding model using data analysis software provided by Pall ForteBio.