Fig. 3

Biolayer interferometry analyses of Bp26 binding to ECM proteins. Heat-inactivated BSA (a), collagen I (b), fibronectin (c), and laminin (d) 521, 10 μg/ml in 10 mM sodium acetate, pH 4 (ForteBio), were respectively coupled onto AR2G sensors (ForteBio) with immobilization levels between 1.5 to 2.0 nm. For kinetics analysis, Bp26 was diluted in the running kinetics buffer (ForteBio) with additional 0.15 M NaCl to reduce the non-specific binding of Bp26 to the reference sensor. The concentrations tested were 0, 125, 250, 500, and 1000 nM. All the experiments were performed at 30 °C, including association for 5 min, and dissociation for 15 min. The raw data were processed by reference subtraction and data correction. E. Biolayer interferometry analyses of vitronectin binding to immobilized Bp26 Bionylated Bp26 was captured onto SA sensors (ForteBio) with immobilization levels of 2.0 nm. Vitronectin was diluted in the running kinetics buffer (ForteBio) to the concentrations of 75, 300, 600, and 1200 nM. All the experiments were performed at 30 °C, including association for 5 min, and dissociation for 15 min. The raw data were processed by reference subtraction and data correction.