New approach to measure protein binding based on a parallel artificial membrane assay and human serum albumin.

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Lazaro E, Lowe PJ, Briand X, Faller B

New approach to measure protein binding based on a parallel artificial membrane assay and human serum albumin.

J Med Chem. 2008 Apr 10;51(7):2009-17. doi: 10.1021/jm7012826. Epub 2008 Mar 19.

PubMed ID

18348514 [ View in PubMed

]

Abstract

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.

DrugBank Data that Cites this Article

Binding Properties

Drug	Target	Property	Measurement	pH	Temperature (°C)
Chlorpromazine	Serum albumin	Kd (nM)	54954	N/A	N/A	Details
Diazepam	Serum albumin	Kd (nM)	24547	N/A	N/A	Details
Diclofenac	Serum albumin	Kd (nM)	2455	N/A	N/A	Details
Diflunisal	Serum albumin	Kd (nM)	1230	N/A	N/A	Details
Tolbutamide	Serum albumin	Kd (nM)	6026	N/A	N/A	Details
Warfarin	Serum albumin	Kd (nM)	4677	N/A	N/A	Details