Self-tuning interfacial architecture for Estradiol detection by surface plasmon resonance biosensor.

This study reports the operation principles for reusable SPR biosensors utilizing nanoscale-specific electrostatic levitation phenomena in their sensitive layer design. Functional macromolecular building blocks localized near the "charged" surface by a variety of weak electrostatic interactions create a flexible and structurally variable architecture. A proof-of-concept is demonstrated by an immunospecific detection of 17ß-Estradiol (E2) following the competitive inhibition format. The sensing interfacial architecture is based on the BSA-E2 conjugate within the BSA matrix immobilized on the "charged" (as a result of guanidine thiocyanate treatment) gold surface at pH 5.0. Kinetic analysis for different E2 concentrations shows that using parameter ß of the stretched exponential function ~(1-exp(-(t/τ)ß) as an analyte-specific response measure allows one to substantially decrease the low detection limit (down to 10-3ng/ml) and increase the dynamic range (10-3-103ng/ml) of the SPR biosensor. Finally, it's concluded that the created interfacial architecture is a typical complex system, where SPR response is formed by the stochastic interactions within the whole variety of processes in the system. The E2 addition destroys the uniformity of the reaction space (where an interaction of the antibody (Ab) and the analog of E2 in the self-tuneable matrix takes place) by the redistribution of the immunospecific complexes Ab(E2)x (x=0, 1, 2) dependent on E2 concentration. Binding dynamics changes are reflected in the values of ß which summarize in compact form all "hidden" information specific for the evolving distributed interfacial system.

A simple SPR-based method for the quantification of the effect of potential virus inhibitors.

Here, we describe a highly sensitive method that allows for the correct quantification of inhibition effect with a higher degree of accuracy directly at the molecular level. The protocol involves two stages, namely serological virus titration in comparison with the same procedure for virus-effector mixture. Owing to the robustness of the analysis this assay can be performed on crude cellular and plant extracts, and therefore it may be suitable for the routine analysis of clinical samples, or in the field. The efficiency of the approach to the quantification of the inhibition effect of polysaccharide glucuronoxylomannan (GXM) on the infection efficiency of the tobacco mosaic virus (TMV) was investigated using advanced serological approaches based on label-free surface plasmon resonance technique. It was shown that GXM drastically decreases the efficiency of TMV infection by blocking up to 70% of the virus shell. The obtained results are in conformity with the method of indicator plant infection.