[Detection of pyrethroids by spectral correlation interferometry].

A label-free method based on spectral correlation interferometry has been developed for highly sensitive detection of pyrethroids by competitive immunoassay on the surface of sensor chips made of widely available microscopy glass cover slips. It is shown that the method allows independent optimization of each step of the sensor surface modification. This fact may be used to increase the efficiency of development of protocols for a wide spectrum of immunoassays that employ glass surface as a solid phase. Detection of 3-phenoxybenzoic acid, which is one of the most stable metabolites of a large number of pyrethroids, on the surface of the optimized sensor chips has been demonstrated on the level of 15 pg/ml. That is 50 times better than the sensitivity of the enzyme-linked immunosorbent assay (ELISA).

Development of immunoassays using interferometric real-time registration of their kinetics.

A method for effective development of solid-phase immunoassays on a glass surface and for optimization of related protocols by highly sensitive quantitative monitoring of each assay step has been proposed and experimentally implemented. The method is based on the spectral correlation interferometry (SCI) that allows real-time measuring of the thickness of a biomolecular layer bound to the recognition molecular receptors on the sensor chip surface. The method is realized with compact 3-channel SCI-biosensors that employ as the sensor chips standard cover glass slips without deposition of any additional films. Different schemes for antibody immobilization on a glass surface have been experimentally compared and optimized toward a higher sorption capacity of the sensor chips. Comparative characterization of the kinetics of each immunoassay stage has been implemented with the optimized protocols: i) covalent immobilization of antibody on an epoxylated surface and ii) biotinylated antibody sorption on a biotinylated surface via a high-affinity biotin-streptavidin bond. We have shown that magnetic nanoparticles employed as labels with model detection of cardiac troponin I further amplify the SCI signal, resulting in 100-fold improvement of the detection limit. The developed protocols can also be used with the alternative immunoassay platforms, including the label methods based on registration of only the final assay result, which is the quantity of bound labels.