LPS-binding activity of DEFB114.
a, recombinant DEFB114 bound on LPS beads was eluted with buffers containing variant concentrations of NaCl and 4 m urea and analyzed on a 15% SDS-polyacrylamide gel. Lane 1, DEFB114 prior to loading on the LPS beads; lane 2, flow-through after loading; lanes 3–6, DEFB114 eluted with NaCl concentrations of 0.1, 0.5, 1.0, and 2.0 m, respectively; lanes 7 and 8, DEFB114 eluted with 4 m urea. b, the binding of DEFB114 to LPS beads was attenuated with competitive LPS. The eluates with 4 m urea were analyzed on a 15% SDS-polyacrylamide gel. Lanes 1 and 2, similar to a; lanes 3–7, eluates of DEFB114 with different concentrations of competitive LPS (0.1, 0.2, 0.4, 0.8, and 1.6 mg/ml, respectively). The association and dissociation of LPS for DEFB114 (c) as well as for alk-DEFB114 (d) was monitored by the Octet interferometer (ForteBio).