ИСТИНА

FLUORESCENCE POLARIZATION IMMUNOASSAYS FOR CARBAMAZEPINE L. Oberleitner1,2, A. Lehmann1, S.A. Eremin3, L.-A. Garbe2, R.J. Schneider1 1BAM Federal Institute for Materials Research and Testing, Berlin, Germany 2Technische Universität Berlin, Berlin, Germany 3M.V.Lomonosov Moscow State University; Moscow, Russia Carbamazepine (5H-dibenz(b,f)azepine-5-carboxamide) is a therapeutic pharmaceutical for the treatment of epilepsy and several mental disorders. Usually the daily intake amounts to 800-1,200 mg. Worldwide approximately 1,014 tons of carbamazepine are consumed per year [1]. Carbamazepine can be detected with immunoassays. These methods are characterized by outstanding selectivity and sensitivity when high-affinity antibodies are available. Enzyme-linked immunosorbent assays (ELISA) for carbamazepine have been developed [2]. This format includes many incubation and washing steps. A faster alternative for the determination of carbamazepine is the fluorescence polarization immunoassay (FPIA). The FPIA method is a competitive immunoassay based on the measurement of fluorescence polarization differences of a reaction mixture of analyte (from sample) competing with an added fluorophore-labeled antigen (tracer) for the binding sites of specific antibodies. This homogenous format avoids the time-consuming steps as for ELISA. Hence, this FPIA format is much faster and is characterized by a very simple handling procedure [3]. The FPIA could be successfully used for detection of pesticides in environmental water samples [4, 5]. Different fluorophore-labeled carbamazepine tracers were synthesized and tested for the suitability for a FPIA for carbamazepine in water samples. Calibration curves for the different labels were set up using the same monoclonal anti-carbamazepine antibody and compared with respect to the sensitivity of the different tracers. Acknowledgement: This research was supported by a Grant of the Russian Foundation for Basic Research 12-03-92105 “Development of microchip-based fluorescence polarization immunoassay of antibiotics in food samples” and by a grant of the German Federal Ministry of Economics and Technology. [1] Y. Zhang, S.-U. Geißen, C. Gal., Chemosphere, 2008, 73, 1151-1161. [2] A. Bahlmann, M. G. Weller, U. Panne, R. J. Schneider, Anal. Bioanal. Chem., 2009, 395, 1809-1820. [3] D. S. Smith and S. A. Eremin, Anal. Bioanal. Chem., 2008, 391, 1499-1507. [4] W-B.. Shim, M. E. Yakovleva, K.-Y. Kim, B.-R. Nam, E. S. Vylegzhanina, A. A. Komarov, S. A. Eremin, and D.-H. Chung, J. Agric. Food Chem., 2009, 57, 791-796. [5] H. Lei, G. Xue, C. Yu, S. A. Haughey, S. A. Eremin, Y. Sun, Z. Wang, Z. Xu, H. Wang, Y. Shen, and Q. Wu, Anal. Methods, 2011, 3, 2334–2340.