Sensitive reagentless electrochemical immunosensor based on an ormosil sol-gel membrane for human chorionic gonadotrophin.

A new organically modified silicate (ormosil) material was synthesized as a matrix to encapsulate enzyme labeled antibody for preparation of immunosensors. The ormosil matrix was prepared by hydrolyzing tetraethyoxysilane and (3-aminopropyl)triethoxysilane in weak alkali solution. It possessed three-dimensional ordered nanoporous structure with high electrical conductivity and good mechanical stability. Its hydrophilicity provided a microenvironment for retaining the biological activity of the immobilized protein. Particularly, using horseradish peroxidase-labeled human chorionic gonadotrophin antibody (HRP-anti-hCG) as a model, the immobilized HRP showed direct electron transfer at about -35 mV with a rate constant of 15.8+/-3.8s(-1). By a simple one-step immunoreaction between human serum chorionic gonadotrophin (hCG) in sample solution and the immobilized HRP-anti-hCG, the differential pulse voltammetric peak current of HRP decreased linearly with an increasing hCG concentration from 0.5 to 50 mIU/ml with a relatively low limit of detection of 0.3 mIU/ml at 3sigma. Excellent analytical performance, fabrication reproducibility and operational stability of the proposed biosensor indicated its promising application in clinical diagnostics.