title: An integrated bioanalytical platform for supporting high-throughput serum protein binding screening.
abstract: Quantification of small molecules using liquid chromatography/tandem mass spectrometry (LC/MS/MS) on a triple quadrupole mass spectrometer has become a common practice in bioanalytical support of in vitro adsorption, distribution, metabolism and excretion (ADME) screening. The bioanalysis process involves primarily three indispensable steps: MS/MS optimization for a large number of new chemical compounds undergoing various screening assays in early drug discovery, high-throughput sample analysis with LC/MS/MS for those chemically diverse compounds using the optimized MS/MS conditions, and post-acquisition data review and reporting. To improve overall efficiency of ADME bioanalysis, an integrated system was proposed featuring an automated and unattended MS/MS optimization, a staggered parallel LC/MS/MS for high-throughput sample analysis, and a sophisticated software tool for LC/MS/MS raw data review as well as biological data calculation and reporting. The integrated platform has been used in bioanalytical support of a serum protein binding screening assay with high speed, high capacity, and good robustness. In this new platform, a unique sample dilution scheme was also introduced. With this dilution design, the total number of analytical samples was reduced; therefore, the total operation time was reduced and the overall throughput was further improved. The performance of the protein binding screening assay was monitored with two controls representing high and low binding properties and an acceptable inter-assay consistency was achieved. This platform has been successfully used for the determination of serum protein binding in multiple species for more than 4000 compounds.
authors.affiliation: Applied Biotechnology, Bristol-Myers Squibb, 5 Research Parkway, Wallingford, CT 06492, USA. jun.zhang1@bms.com
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.forename: Wilson Z
authors.initials: WZ
authors.lastname: Shou
authors.forename: Marianne
authors.initials: M
authors.lastname: Vath
authors.forename: Kasia
authors.initials: K
authors.lastname: Kieltyka
authors.forename: Jennifer
authors.initials: J
authors.lastname: Maloney
authors.forename: Larry
authors.initials: L
authors.lastname: Elvebak
authors.forename: Jeremy
authors.initials: J
authors.lastname: Stewart
authors.forename: John
authors.initials: J
authors.lastname: Herbst
authors.forename: Harold N
authors.initials: HN
authors.lastname: Weller
mesh_terms: D000818:Animals; D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism*; D002853:Chromatography, Liquid; D004285:Dogs; D055808:Drug Discovery / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D016014:Linear Models; D008252:Macaca fascicularis; D008297:Male; D051379:Mice; D010599:Pharmacokinetics; D011485:Protein Binding; D051381:Rats; D015203:Reproducibility of Results; D018511:Systems Integration; D053719:Tandem Mass Spectrometry
chemical_list: D001798:Blood Proteins