A differential pulse voltammetric immunoassay associated with the TMB-H2O2-HRP for the determination of IgE antibody was developed. The enzymatic reaction product produced by HRP catalyzing the oxidation of H2O2 with ...A differential pulse voltammetric immunoassay associated with the TMB-H2O2-HRP for the determination of IgE antibody was developed. The enzymatic reaction product produced by HRP catalyzing the oxidation of H2O2 with TMB exhibits a sensitive voltammet-ric response at 0.10V(vs.Ag/AgCl) in pH4.0 phosphate-citrate buffer solution. The de-tection limit of the present method for IgE antibody was 4.0IU/mL, which was four times lower than that obtained by traditional spectrophotometric ELISA procedure. It has been ap-plied to the determination of IgE in human serum with satisfactory results.展开更多
With 3, 3’5, 5’-tetramethylbenzidine（TMB） as the detection substrate, a reliable and highly selective method was established and optimized for the determination of Lactoperoxidase（LP） activity in raw milk. The m...With 3, 3’5, 5’-tetramethylbenzidine（TMB） as the detection substrate, a reliable and highly selective method was established and optimized for the determination of Lactoperoxidase（LP） activity in raw milk. The method was based on the enzymatic reaction principle, where hydrogen peroxide oxidated TMB in the presence of LP. The optimized conditions of this assay system were obtained, consisting of 20 mmol · L-1 TMB solution, 0.6 mmol · L-1 hydrogen peroxide and 0.1 mol · L-1 Citric Acid（CA）/0.2 mol · L-1 disodium hydrogen phosphate（Na P） buffer（pH 4.8）. TMB detection method was applied to the analysis of LP in milk samples with a practical working concentration range from 2 to 14 mg · L-1. The intra-and inter-batch variation coefficients were all below 5%, indicating a good repeatability. Confirmation test between TMB method and 2, 2-azinobi（3-ethylbenzothiazoline-6-sulphonate） diammonium salt（ABTS） method was carried out, and the results of TMB assay were in accordance with that of ABTS method.展开更多
Traditional methods for detecting lactoperoxidase （LP） are complex and time-consuming, so a test strip was made based on the enzymatic reaction principle to enable quick and convenient detection of LP in raw milk. I...Traditional methods for detecting lactoperoxidase （LP） are complex and time-consuming, so a test strip was made based on the enzymatic reaction principle to enable quick and convenient detection of LP in raw milk. In this study 0.1 mol/L citric acid （CA）/0.2 mol/L disodium hydrogen phosphate （NAP） buffer solution （pH 5.0）, 22 mmol/L 3,3＇,5,5＇-tetramethylbenzidine （TMB）, 0.6 mmol/L hydrogen peroxide （H202）, and 0.5% Tween-20 or 0.3% cetyltri- methyl ammonium bromide （CTAB） were optimal for preparing a quick, sensitive, and accurate LP test strip. The coefficient of variation （CV） of the estimated LP concentrations ranged from 2.47% to 6.72% and the minimum LP concentration detected by the test strip was 1-2 mg/L. Estimates of active LP in sixteen raw milk samples obtained using the test strip or the TMB method showed a good correlation （c=0.9776）. So the test strip provides a quick, convenient, and accurate method for detecting the LP concentration of raw milk.展开更多
文摘A differential pulse voltammetric immunoassay associated with the TMB-H2O2-HRP for the determination of IgE antibody was developed. The enzymatic reaction product produced by HRP catalyzing the oxidation of H2O2 with TMB exhibits a sensitive voltammet-ric response at 0.10V(vs.Ag/AgCl) in pH4.0 phosphate-citrate buffer solution. The de-tection limit of the present method for IgE antibody was 4.0IU/mL, which was four times lower than that obtained by traditional spectrophotometric ELISA procedure. It has been ap-plied to the determination of IgE in human serum with satisfactory results.
基金Supported by Project for Research and Development of Harbin Aapplication Technology(2016RAQXJ046)the National "Twelfth Five-year" Plan for Science and Technology Support Program of China(2013BAD18B06)
文摘With 3, 3’5, 5’-tetramethylbenzidine（TMB） as the detection substrate, a reliable and highly selective method was established and optimized for the determination of Lactoperoxidase（LP） activity in raw milk. The method was based on the enzymatic reaction principle, where hydrogen peroxide oxidated TMB in the presence of LP. The optimized conditions of this assay system were obtained, consisting of 20 mmol · L-1 TMB solution, 0.6 mmol · L-1 hydrogen peroxide and 0.1 mol · L-1 Citric Acid（CA）/0.2 mol · L-1 disodium hydrogen phosphate（Na P） buffer（pH 4.8）. TMB detection method was applied to the analysis of LP in milk samples with a practical working concentration range from 2 to 14 mg · L-1. The intra-and inter-batch variation coefficients were all below 5%, indicating a good repeatability. Confirmation test between TMB method and 2, 2-azinobi（3-ethylbenzothiazoline-6-sulphonate） diammonium salt（ABTS） method was carried out, and the results of TMB assay were in accordance with that of ABTS method.
基金supported by the National "Twelfth Five-Year" Plan for Science and Technology Support Program of China(No.2013BAD18B06)the Open Research Fund for the Key Laboratory of Dairy Science of Northeast Agricultural University(No.KLDS201201)the Innovative Research Team of Higher Education of Heilongjiang Province(No.2010td11),China
文摘Traditional methods for detecting lactoperoxidase （LP） are complex and time-consuming, so a test strip was made based on the enzymatic reaction principle to enable quick and convenient detection of LP in raw milk. In this study 0.1 mol/L citric acid （CA）/0.2 mol/L disodium hydrogen phosphate （NAP） buffer solution （pH 5.0）, 22 mmol/L 3,3＇,5,5＇-tetramethylbenzidine （TMB）, 0.6 mmol/L hydrogen peroxide （H202）, and 0.5% Tween-20 or 0.3% cetyltri- methyl ammonium bromide （CTAB） were optimal for preparing a quick, sensitive, and accurate LP test strip. The coefficient of variation （CV） of the estimated LP concentrations ranged from 2.47% to 6.72% and the minimum LP concentration detected by the test strip was 1-2 mg/L. Estimates of active LP in sixteen raw milk samples obtained using the test strip or the TMB method showed a good correlation （c=0.9776）. So the test strip provides a quick, convenient, and accurate method for detecting the LP concentration of raw milk.
