title: Automation of plasma protein binding assay using rapid equilibrium dialysis device and Tecan workstation.
abstract: Binding of drug molecules to plasma proteins is an important parameter in assessing drug ADME properties. Plasma protein binding (PPB) assays are routinely performed during drug discovery and development. A fully automated PPB assay was developed using rapid equilibrium dialysis (RED) device and Tecan workstation coupled to an automated incubator. The PPB assay was carried out in unsealed RED plates which allowed the assay to be fully automated. The plasma pH was maintained at 7.4 during the 6-h dialysis under 2% CO2condition. The samples were extracted with acetonitrile and analyzed by liquid chromatography tandem mass spectrometry. The percent bound results of 10 commercial drugs in plasma protein binding were very similar between the automated and manual assays, and were comparable to literature values. The automated assay increases laboratory productivity and is applicable to high-throughput screening of drug protein binding in drug discovery.
authors.affiliation: Drug Metabolism and Pharmacokinetics, Vertex Pharmaceuticals, Inc. 50 Northern Ave, Boston, MA 02210, USA. Electronic address: zhengqi_ye@vrtx.com.
authors.forename: Zhengqi
authors.initials: Z
authors.lastname: Ye
authors.affiliation: Drug Metabolism and Pharmacokinetics, Vertex Pharmaceuticals, Inc. 50 Northern Ave, Boston, MA 02210, USA.
authors.forename: Craig
authors.initials: C
authors.lastname: Zetterberg
authors.affiliation: Drug Metabolism and Pharmacokinetics, Vertex Pharmaceuticals, Inc. 50 Northern Ave, Boston, MA 02210, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Gao
mesh_terms: D001331:Automation; D001798:Blood Proteins / Q000032:analysis*; D002853:Chromatography, Liquid; D003956:Dialysis; D011485:Protein Binding; D006435:Renal Dialysis
chemical_list: D001798:Blood Proteins
keywords: ADME; High-throughput plasma protein binding assay; RED device; Tecan workstation