New approach to measure protein binding based on a parallel artificial membrane assay and human serum albumin.
We report here a new, label-free approach to measure serum protein binding constants. The assay is able to measure HSA K d values in the milli-molar to micromolar range. The protein is not immobilized on any surface and the assay self-corrects for nonspecific adsorption. No mass balance is required to get accurate binding constants and it is not necessary to wait for equilibrium to extract the binding constant. The assay runs in a 96-well format using commercially available parts and is, therefore, relatively easy to implement and automate. As the chemical membranes used are not water permeable, there is no volume change due to the osmotic pressure and pretreatment (soaking) is not necessary. The concept can potentially be extended to other proteins and could thus serve as a label-free technique for general binding constant measurements.
Lázaro
Elisabet
E
Departament de Química Analítica, Universitat de Barcelona, Martí i Franquès 1, Barcelona, Spain.
Lowe
Philip J
PJ
Briand
Xavier
X
Faller
Bernard
B
D006801:Humans; D007700:Kinetics; D008567:Membranes, Artificial; D010539:Permeability; D011485:Protein Binding; D012709:Serum Albumin / Q000737:chemistry*
D008567:Membranes, Artificial; D012709:Serum Albumin