An integrated bioanalytical platform for supporting high-throughput serum protein binding screening
Article first published online: 15 NOV 2010
Copyright © 2010 John Wiley & Sons, Ltd.
Rapid Communications in Mass Spectrometry
Volume 24, Issue 24, pages 3593–3601, 30 December 2010
How to Cite
Zhang, J., Shou, W. Z., Vath, M., Kieltyka, K., Maloney, J., Elvebak, L., Stewart, J., Herbst, J. and Weller, H. N. (2010), An integrated bioanalytical platform for supporting high-throughput serum protein binding screening. Rapid Commun. Mass Spectrom., 24: 3593–3601. doi: 10.1002/rcm.4817
- Issue published online: 12 NOV 2010
- Article first published online: 15 NOV 2010
- Manuscript Accepted: 10 OCT 2010
- Manuscript Revised: 8 OCT 2010
- Manuscript Received: 5 AUG 2010
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. Copyright © 2010 John Wiley & Sons, Ltd.