immunosensors;
ormosil;
sol-gel;
enzyme labeled antibody;
human chorionic gonadotrophin;
D O I:
10.1016/j.bios.2006.12.010
中图分类号:
Q6 [生物物理学];
学科分类号:
071011 ;
摘要:
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 tetraethyoxysi lane and (3-aminopropyl)triethoxysitane 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 +/- 3.8 s(-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 voltarnmetric peak current of HRP decreased linearly with an increasing hCG concentration from 0.5 to 50 mIU/mI with a relatively low limit of detection of 0.3 mIU/mI at 3 sigma. Excellent analytical performance, fabrication reproducibility and operational stability of the proposed biosensor indicated its promising application in clinical diagnostics. (c) 2006 Elsevier B.V. All rights reserved.