ИСТИНА

The qualitative and quantitative detection of biologically active compounds must be made for any kind of research and application. Development of reliable analytical methodologies and instruments for screening biologically active compounds should be of high priority. Moreover, the screening method could be for the simultaneous detection of several substances with high-throughput screening (HTS). The current methods of assays like chromatography techniques and enzyme-linked immunosorbent assay (ELISA) have several limitations for HTS. The analytical methods applied for detection of biologically active compounds must ideally be quick, cheap, and simple with high throughput in a wide operating range. These requirements are best met by Fluorescence Polarization Immunoassay (FPIA). FPIA as any immunoassay have several advantages in sensitivity and specificity as well as a useful alternative approach to ELISA because it is a homogeneous method without any separation and washing steps. FPIA is direct competitive immunoassay method, which based on the measurement of fluorescence polarization (FP) value for reaction mixture of sample and immunoreagents: fluorescent-labeled antigen (tracer) and specific antibodies. To performance the assay only the tracer and the antibody solutions must be added to aliquot of liquid sample (typically 10-50 µl), mixed and measured. The total time for an assay is few seconds or minutes. Recent results for preparation of immunoreagents (tracers with different structures and specific antibodies) and development of FPIA for several biologically active compounds like Chloramphenicol, Sulfamethazine, Fluoroquinolones will be presented and discussed. The limit of detection for compounds could be up to 10 ng/ml. These immunoreagents and developed FPIA could be adapted for detection of biologically active compounds in flow system using flow-cell detector for Fluorescence Polarization. Acknowledgements: The research was supported by cooperation Japan and Russia (RFBR, project 12-03-92105).