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21.
A simple direct enzyme immunoassay for semiquantitative detec tion of antibodies is suggested. It is based on the difference
in diffusion rates in a gel for a synthetic low-mol-wt antigen and of its complexes with antibodies to be detected. Sensitivity
and specificity of the devel oped assay are equal to an ELISA method. The assay has been tested with antibodies against HIV
protein gp41 in rabbit serum. Possible applications and limitations of the method are discussed. 相似文献
22.
23.
Bulk-modified epoxy-graphite tyrosinase biosensors were fabricated by four different procedures. The influence of these fabrication procedures on the analytical performance of the enzyme electrode in an amperometric wall-jet flow cell has been studied. The bioprobe performance is assessed by cyclic voltammetry. Higher current densities and narrower peaks were obtained when the enzyme was introduced in the dry state into the epoxy-graphite material, instead of introducing it previously dissolved in the buffer. In the F1 system responses of 11.79 μA cm−2 and 1.43 μA cm−2 are then obtained for catechol and phenol respectively for 50 μL injections of 20 μM solutions. Moreover, if gold/palladium is introduced into the epoxy-graphite, a further increase in current is achieved resulting in 27.70μA cm−2 and 4.90μA cm−2for catechol and phenol, respectively. This biosensor can operate in aqueous as well as in mixed aqueous-organic environments. 相似文献
24.
Emmanuel Burgos 《Tetrahedron letters》2004,45(17):3465-3469
4-deoxy-4-Phosphonomethyl-d-erythronate, an isosteric and hydrolytically stable analogue of the known ribose-5-phosphate isomerase inhibitor 4-deoxy-4-phospho-d-erythronate, was obtained by a 14-step synthesis from d-arabinose through an highly improved synthesis of the precursor 5-deoxy-5-phosphonomethyl-d-arabinose. The title compound appears as the first stable and potent competitive inhibitor of the enzyme catalyzed isomerization of ribose-5-phosphate to d-ribulose-5-phosphate (Ki=74 μM, Km/Ki=100), exhibiting only a 3-fold weaker inhibitory activity than its phosphate analogue. 相似文献
25.
Adam Bange Jian Tu Xiaoshan Zhu Chong Ahn H. Brian Halsall William R. Heineman 《Electroanalysis》2007,19(21):2202-2207
A microbead based sandwich immunoassay for MS2 bacteriophage was developed using an interdigitated array (IDA) electrode with nanoscale dimensions (220 nm electrode width, 620 nm gap). The IDA was fabricated using an electron beam lithographic lift‐off technique. After an antibody‐assisted capture of MS2 using paramagnetic microbeads, a β‐galactosidase labeled secondary antibody was used to convert p‐aminophenyl‐β‐D ‐galactopyranoside (PAPG) into the redox active p‐aminophenol (PAP). Amperometric detection of PAP with IDA electrodes at +300 and ?200 mV vs. a Ag/AgCl reference electrode was used to measure the result, detecting MS2 concentrations as low as 10 ng/mL. 相似文献
26.
Enzyme heterobilayer-modified electrodes were fabricated by successively covalently binding to the surface of a tin(IV) oxide plate horseradish peroxidase (HRP), then an oxidase (lactate, pyruvate or cholesterol oxidase or uricase), which liberates hydrogen peroxide by reaction with the respective substrate. The cooperative action of oxidase-HRP leads to an efficient amperometric sensor system with the minimum amount of enzyme immobilized on an electrode. 相似文献
27.
The construction and response of an immobilized enzyme modified electrode as an amperometric sensor is described. Xanthine oxidase was adsorbed on a carbon paste electrode and physically entrapped with a semipermeable membrane. Uric acid, the product of the enzymatic reaction, was oxidized electrochemically at +0.4 V vs. Ag/AgCl, yielding a steady-state current directly related to the bulk concentration of the substrate. Hypoxanthine and xanthine were determined in the range 5–100 μM at Ph 7.2 with good precision. Interferences are discussed. 相似文献
28.
Clinical applications of chemiluminescence 总被引:8,自引:0,他引:8
This article reviews the clinical applications of chemiluminescence in routine testing and surveys the diverse applications of chemiluminescence in clinical research. In routine clinical testing, chemiluminescent labels (acridinium ester, acridinium sulfonamide) and detection reactions for peroxidase and alkaline phosphatase enzyme labels (luminol and adamantyl 1,2-dioxetane-based reactions, respectively) are widely used in immunoassay and nucleic acid probe assays (e.g. hybridization protection assay, Hybrid Capture® assay). In clinical research the sensitivity, dynamic range and diversity of chemiluminescent assays has led to a vast range of applications, notably in protein and nucleic acid blotting, microarray-based assays, monitoring reactive oxygen species, and as detection reactions for substances separated by HPLC, capillary electrophoresis (CE), and flow-injection analysis. 相似文献
29.
Yannis Dotsikas 《Analytica chimica acta》2004,509(1):103-109
This study describes the employment of a novel imidazole-substituted phenol [4-(1-imidazolyl)phenol] as a highly potent signal enhancer in a horseradish peroxidase (HRP)-luminol chemiluminescence (CL) immunoassay. This competitive-type immunoassay for the model antigen fentanyl is based on the use of fentanyl polyclonal antibody immobilized on white microtiter plates and a biotinylated bovine serum albumin (BSA)-fentanyl derivative as a tracer. The latter was detected by means of streptavidin labeled with HRP, resulting in the generation of a high-intensity and relatively stable chemiluminescent signal, immediately after the addition of the substrate solution (NOAS). The developed method fulfilled the requirements of accuracy (percentage recovery ranged from 93.8 to 107%) and precision (intra- and inter-assay CVs were 2.5-5.2 and 4.5-11.9%, respectively). Its plasma detection limit (1.05 pg ml−1) was lower than those of previous immunoassays. The novel assay was compared in terms of sensitivity and concentration range with other common HRP substrate systems: luminol-p-iodophenol-H2O2 and TMB-H2O2. Finally, the described method was compared with an HRP-fentanyl conjugate-based assay, similar to commercially available kits (SKIT), employing the novel substrate solution for both assays and the differences observed were explained by applying previously described models. The detection limit was 4.82 pg ml−1 for SKIT, recovery values were 94.2-105% and intra- and inter-assay CVs were 2.5-5.2 and 4.5-11.9%, respectively. In conclusion, the proposed assay could be utilized for a wide range of molecules and replace the existing enzyme-labeled antigen-based kits. 相似文献
30.