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1.
Biosens Bioelectron ; 90: 91-95, 2017 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-27886605

RESUMO

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.


Assuntos
Técnicas Biossensoriais , Estradiol/isolamento & purificação , Ressonância de Plasmônio de Superfície , Estradiol/química , Estradiol/imunologia , Ouro/química , Limite de Detecção , Soroalbumina Bovina/química , Soroalbumina Bovina/imunologia
2.
Analyst ; 140(9): 3225-32, 2015 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-25763411

RESUMO

An ultra-sensitive gas phase biosensor/tracer/bio-sniffer is an emerging technology platform designed to provide real-time information on air-borne analytes, or those in liquids, through classical headspace analysis. The desired bio-sniffer measures gaseous 17α- ethinylestradiol (ETED) as frequency changes on a quartz crystal microbalance (QCM), which is a result of the interactions of liquid sample components in the headspace (ETED and water) with a biorecognition layer. The latter was constructed by immobilization of polyclonal antiserum against a phenolic A-ring of estrogenic receptors through protein A. The QCM response exhibited stretched exponential kinetics of negative frequency shifts with reversible and "irreversible" components of mass uptake onto the sensor surface in static headspace conditions when exposed to water solutions of ETED over the sensor working range, from 10(-10) to 10(-17) g L(-1). It was shown that the variations in the QCM response characteristics are due to the change of the water-binding capacity of the sensing layer induced by protein transformations initiated by the binding of ETED molecules. This result is well correlated with the natural physiological function of estrogens in controlling the homeostasis of body fluids in living beings.


Assuntos
Técnicas Biossensoriais/instrumentação , Etinilestradiol/análise , Técnicas de Microbalança de Cristal de Quartzo/instrumentação , Adsorção , Animais , Desenho de Equipamento , Ligação Proteica , Coelhos , Proteína Estafilocócica A/metabolismo , Água/química
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