title: Study of binding constant of toll-like receptor 4 and lipopolysaccharide using capillary zone electrophoresis.
abstract: This short communication describes the interaction between toll-like receptor 4 (TLR4) and lipopolysaccharide (LPS, its specific ligand) using analytical methods. Their interaction has been evidenced in many reports. Nevertheless, there are few reports focused on their binding constant. In this research, the interaction between TLR4 and LPS is investigated using mobility shift method by CZE. To optimize the electrophoresis conditions, the effecting factors, running buffer, sample concentration, injection duration, and operation voltage of electrophoretic on the mobility shift are studied in detail. Electrophoresis conditions were described as follows: borate buffer (pH 7.4, 20 mM), 5 s for 50 mbar pressure injection duration, and 13 kV of separation voltage in 41.5 cm fused silica capillary with 75 μm id and 375 μm od. The combination constant of TLR4 and LPS is calculated using Scatchard methods. The Scatchard liner correlation is y=-0.0165x+0.1456, binding constant is K=1.65 x 10⁴  (g/mL)⁻¹.
authors.affiliation: School of Life Science, Beijing Institute of Technology, Beijing, P. R. China.
authors.forename: Huiqing
authors.initials: H
authors.lastname: Fu
authors.forename: Jimin
authors.initials: J
authors.lastname: Li
authors.forename: Weiwei
authors.initials: W
authors.lastname: Meng
authors.forename: Runan
authors.initials: R
authors.lastname: Dong
authors.forename: Rongji
authors.initials: R
authors.lastname: Dai
authors.forename: Yulin
authors.initials: Y
authors.lastname: Deng
mesh_terms: D019075:Electrophoresis, Capillary / Q000379:methods*; D016014:Linear Models; D008070:Lipopolysaccharides / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D051197:Toll-Like Receptor 4 / Q000737:chemistry / Q000378:metabolism*
chemical_list: D008070:Lipopolysaccharides; D051197:Toll-Like Receptor 4