A highly sensitive, fast and stable conductometric immunosensor for determination of interleukin-6 (IL6) in humans is developed by encapsulation of horseradish peroxidase-labeled interleukin-6 antibody (HRP-anti-IL6) in poly(amidoamine) fourth-generation dendrimer (dendrimer) and colloidal gold (nanogold) modified composite architecture. The presences of nanogold and dendrimer provided a congenial microenvironment for the immobilized biomolecules and decreased the electron transfer impedance, leading to a direct electrochemical behavior of the immobilized HRP. The formation of the antibody-antigen complex by a simple one-step immunoreaction between the immobilized HRP-anti-IL6 and IL6 in sample solution introduced a barrier of direct electrical communication between the immobilized HRP and the gold electrode surface, thus local conductivity variations could be detected by the HRP electrocatalytic reaction in 0.02 M phosphate buffer solution (pH 7.0) containing 50 mM H 2 O 2 , 0.01 M KI and 0.15 M NaC1. Under optimal conditions, the proposed immunosensor exhibited a good conductometric response to IL6 in a linear range from 30 to 300 pg/mL with a relatively low detection limit of 10 pg/mL at 3d. The precision and reproducibility are acceptable with the intra-assay CV of 7.3% and 5.6% at 100 and 200 pg/mL IL6, respectively. The storage stability of the proposed immunosensor is acceptable in a pH 7.0 PBS at 4 8C for 8 days. Importantly, the proposed methodology could be extended to the detection of other antigens or biocompounds.
Various sensor-based immunoassay methods have been extensively developed for the detection of interleukin-6 (IL6), but most often exhibit low detection signals and low detection sensitivity, and are unsuitable for routine use. The aim of this work is to develop a simple and sensitive conductometric immunoassay for IL6 in human serum by using an organic/inorganic hybrid membrane-functionalized interface. Initially, thionine-bound 3,4,9,10-perylenetetracarboxylic acid was doped into colloidal alumina, then nanogold particles were immobilized onto the thionine surface, and then horseradish peroxidase-labeled anti-IL6 antibodies were conjugated on the nanogold surface. The organic/inorganic hybrid membrane provides a good microenvironment for the immobilization of biomolecules, enhanced the surface coverage of protein, and improved the sensitivity of the immunosensor. The performance and factors influencing the performance of the immunosensor were evaluated. The detection is based on the change in local conductivity before and after the antigen-antibody interaction in 0.02 M phosphate buffer solution (pH 6.8) containing 50 microM H(2)O(2), 0.01 M KI and 0.15 M NaC1. Under optimal conditions, the proposed immunosensor exhibited a wide linear range from 25 to 400 pg/ml towards IL6 with a relatively low detection limit of 5 pg/ml (S/N = 3). The stability, reproducibility and precision of the immunosensor were acceptable. 37 serum specimens were assayed by the developed immunosensor and standard enzyme-linked immunosorbent assay, respectively, and the results obtained were almost consistent. More importantly, the detection methodology provides a promising approach for other proteins or biosecurity.
Immunophenotyping evaluation is of particular importance for the clinical diagnosis, therapy, and prognosis of viral hepatitis. In this study, an integrated micro flow device has been developed to detect the differentiated antigens/antibodies for immunophenotyping of viral hepatitis. The sensors were fabricated with plasma-polymerized ethylenediamine film (PPF) and nanometer-sized gold particles (nanogold) on which the different hepatitis B antigens/antibodies (markers) were subsequently immobilized. Monitoring the changes in the potential signals before and after the antigen-antibody interaction provides the basis for an immunoassay that is simple, rapid, and cost-effective. It permits the detection of hepatitis B in the dynamic concentration range of 2 orders of magnitude (10(-6) g x L(-1) - 10(-4) g x L(-1)). Up to 7 successive assay cycles with retentive sensitivity were achieved for the sensors regenerated by 8 M urea. Moreover, the microfluidic device was applied to evaluate a number of practical specimens with analytical results in acceptable agreement with those clinically classified. The newly proposed multiparameter analysis technique provides a feasible alternative tool for the diagnosis and monitoring of hepatitis B.
An efficient biosensing substrate based on ZrO 2 / DNA-derivated polyion complex (PIC) membrane has been developed for the determination of hydrogen peroxide (H 2 O 2 ) in this study. To fabricate such a PIC membrane, ZrO 2 nanoparticles were initially electrodeposited on the bare gold electrode (ZrO 2 /Au), and deoxyribonucleic acid (DNA)-doped hemoglobin mixture was then assembled onto the ZrO 2 /Au surface. The double-strand DNA provided a biocompatible microenvironment for the immobilization of biomolecules, greatly amplified the surface coverage of biomolecules on the electrode surface, and improved the sensitivity of the biosensor. The fabricated procedure of the proposed biosensor was characterized by cyclic voltammetry, electrochemical impedance spectroscopy, and atomic force microscopy. The performance and factors influencing the performance of the biosensor were also evaluated. Under optimal conditions, the developed biosensor exhibited a well-defined electrochemical behavior toward the reduction of H 2 O 2 ranging from 1.1 μM to 2.3 mM with a detection limit of 0.5 μM (S/N=3). The biosensor was applied to the determination of H 2 O 2 in milk with satisfactory results. It is important to note that the PIC membrane provided an alternative substrate for the immobilization of other proteins.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.