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Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells
Published on: May 22, 2014
Inmunosensor de reconocimiento externo de sonda interna fabricado en nano canales para la detección electroquímica
Huanrui Wang1,2, Hongxin Wang3, Fei Yan3
1Department of Urology, Peking University People's Hospital Beijing 100044 China.
Abstract:
Highly sensitive and selective detection of interleukin-1β (IL-1β) enables dynamic monitoring of IL-1β levels in complex samples, providing significant clinical value for the prevention of early pathogenesis of urosepsis. Herein, a dual-functional "internal probe-external recognition" immunosensor was developed on nanochannel array, spatially separating signal generation from target recognition for label-free detection of IL-1β. A polydopamine (PDA) adhesive layer was utilized to stabilize the growth of amino-functionalized vertically-ordered mesoporous silica film (NH2-VMSF) on a glassy carbon electrode (GCE). Using the dual functionality of NH2-VMSF, gold nanoparticles (AuNPs) were electrochemically deposited within the NH2-VMSF nanochannels as immobilized electrochemical probes, while antibodies were covalently immobilized on the outer surface of NH2-VMSF to act as biological recognition elements. Upon binding with IL-1β, the immunosensor specifically recognized and captured the target, resulting in an increase in interface resistance and a corresponding decrease in the electrochemical signal of AuNPs. This mechanism enabled the quantitative analysis of IL-1β, with a linear detection range from 1 pg mL-1 to 100 ng mL-1 and a limit of detection (LOD) of 0.6 pg mL-1. The immunosensor was successfully applied to IL-1β detection in fetal bovine serum (FBS). By eliminating the need for additional signal probes, the developed immunosensor simplifies the detection process, demonstrates excellent anti-interference and stability, and offers a new approach for reagent-free biomarker detection in complex samples.

