Analytical and Bioanalytical Chemistry

, Volume 388, Issue 5–6, pp 1147–1156 | Cite as

Determination of drug–serum protein interactions via fluorescence polarization measurements

  • Ulf Mathias
  • Manfred Jung
Original Paper


New fast methods for the determination of pharmacokinetic behaviour of potential drug candidates are receiving increasing interest. We present a new homogeneous method for the determination of drug binding and drug competition for human serum albumin and α1-acid glycoprotein that is amenable to high-throughput-screening. It is based on selective fluorescent probes and the measurement of fluorescence polarization. This leads to decreased interference with fluorescent drugs as compared with previously published methods based on similar probes and the measurement of fluorescence intensity. The binding of highly fluorescent drugs that still interfere with the probes can be measured by simply titrating the drugs in a two-component system with the serum protein. The assay may also be used to discover strongly binding protein ligands that are interesting for drug-targeting strategies. Additionally, binding data could be obtained from larger libraries of compounds for in silico predictive pharmacokinetics.


Fluorescence polarization displacement titration of dansylsarcosine (3D-structure as insert) bound to human serum albumin (HSA) by naproxene


Human serum albumin α1-acid glycoprotein High-throughput-screening Early-ADME tests Fluorescence polarization 



We thank Roche Diagnostics (Penzberg, Germany) for financial support, L. Stecher and S. Wittich for exploratory experiments and W. Sippl (Halle) for the 3D graphic for the online edition.

Supplementary material


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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  1. 1.Institute of Pharmaceutical SciencesUniversity of FreiburgFreiburgGermany

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