Enzyme Immunoasay

Monroe, Dan

doi:10.1021/ac00272a719

Cited by 7 publications

(6 citation statements)

References 4 publications

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“…Typically, specific interactions between the biological molecules are studied using a number of commonly known physical and biochemical methods, such as surface plasmon resonance (SPR) [1], enzyme linked immunosorbent assay (ELISA) [2], analytical affinity chromatography (HPLAC) [3], affinity capillary electrophoresis (ACE) [4], etc. Some methods determine only the affinity of studied species, while others also provide information about the kinetics of the interaction.…”

Section: Introductionmentioning

confidence: 99%

Atomic Force Microscopy and Quartz Crystal Microbalance Study of the Lectin-Carbohydrate Interaction Kinetics

et al. 2007

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Two analytical methods, atomic force microscopy and quartz crystal microbalance, were applied to the study of the reaction kinetics occurring between concanavalin A and carboxypeptidase Y, presenting the specific lectin-carbohydrate recognition. The dissociation rate constants for concanavalin A-carboxypeptidase Y complex obtained using both atomic force microscopy and quartz crystal microbalance were of the same order of magnitude: k diss = 0.170 ± 0.060 s −1 and k diss = 0.095 ± 0.002 s −1 , respectively. In addition, each method alone aided in determining other parameters characterizing the studied interaction. Quartz crystal microbalance permitted us to estimate the association rate (k ass = (5.6 ± 0.1) × 10 4 M −1 s −1 ) and the equilibrium (Ka = (0.59 ± 0.01) × 10 6 M −1 ) constants for the binding process occurring between concanavalin A and mannose residues of carboxypeptidase Y under given experimental conditions. Atomic force microscopy in force spectroscopy mode enabled the determination of the energy barrier position of r = 2.29 ± 0.04Å characterizing the dissociation of concanavalin Acarboxypeptidase Y molecular complex. The presented results show that both atomic force microscopy and quartz crystal microbalance can be used to determine quantitative parameters characterizing the specific molecular interaction. Both methods can be easily combined for complementary and/or alternative studies of a chosen molecular interaction. By preparing the samples in the same manner the direct comparison between the data obtained via atomic force microscopy and quartz crystal microbalance can be made.

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Section: Introductionmentioning

confidence: 99%

Atomic Force Microscopy and Quartz Crystal Microbalance Study of the Lectin-Carbohydrate Interaction Kinetics

et al. 2007

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“…In order to reduce the specific binding caused by linking arm, the diazotization method and the glutaraldehyde method were used to couple the terminal amino group on DBP hapten to the carrier proteins for preparing immunogen and coating antigen, respectively. UV-Vis spectrophotometer was used to identify all conjugates, and then the coupling ratios were estimated based on mole absorbance ε and calculated using the following equation (Monroe 1984;Zhang et al 2006a, b):…”

Section: Preparation Of Immunogen and Coating Antigenmentioning

confidence: 99%

Biotin-Streptavidin Enzyme-linked Immunosorbent Assay for the Determination of Dibutyl Phthalate in Beverages and Drinking Water Using a Specific Polyclonal Antibody

Sun

Zhuang

2015

Food Anal. Methods

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A specific polyclonal antibody targeting dibutyl phthalate (DBP) was obtained, and a sensitive indirect competitive biotin-streptavidin enzyme-linked immunosorbent assay (BA-ELISA) was developed for the determination of DBP in beverages and drinking water. Under optimal conditions, good linearity was achieved within a range of 0.02 to 8.97 μg·L −1 . The limit of detection (IC 10 ) was 5 ng·L −1 , and the median inhibitory concentration (IC 50 ) was 0.36 μg·L −1 . The BA-ELISAwas highly selective, with lower cross-reactivity values with DBP analogues (lower than 4 %). Finally, the concentrations of DBP in beverages and drinking water by this method ranged from 0.45 to 7.06 μg·L −1 . Satisfactory recoveries (89.5 to 109.5 %) and variation coefficient values (6.0 to 8.7 %) were obtained. The consistency between the results obtained from BA-ELISA and gas chromatography-mass spectrometry (GC-MS) was 98.8 %, which further confirmed that this proposed BA-ELISA immunoassay was reliable, rapid, sensitive, and accurate for monitoring DBP in the environmental samples.

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“…[1][2][3][4] Enzyme-linked competitive binding methods are based on a competition between the unlabeled analyte and an enzyme-analyte conjugate for a limited number of binding sites. Such methods can be classified as either heterogeneous (solid-phase) or homogeneous.…”

mentioning

confidence: 99%

Enzyme-Linked Competitive Binding Assays Based on the Biotin/Avidin Interaction

et al. 1999

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The strong interaction between biotin and avidin is utilized for the development of enzyme-linked competitive binding assays not only for biotin itself, but also for other biomolecules. Alkaline phosphatase, a single substrate enzyme typically used for heterogeneous types of competitive binding assays, is employed also for homogeneous types. For the biotin assay, the heterogeneous protocol offers a much improved detection capability when compared to the homogeneous type. A simple approach of simulating dose-response curves is introduced. An effective analyte concentration attached to an enzyme label is determined by using the same binder, but with a different enzyme label. The analyte system adapted to the biotin/avidin-mediated homogeneous protocol is digoxin and a monoclonal anti-digoxin antibody. The detection capabilities of these assays, particularly the homogeneous types, are shown to be limited by the detectability of the enzyme conjugate employed.

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Enzyme Immunoasay

Cited by 7 publications

References 4 publications

Atomic Force Microscopy and Quartz Crystal Microbalance Study of the Lectin-Carbohydrate Interaction Kinetics

Atomic Force Microscopy and Quartz Crystal Microbalance Study of the Lectin-Carbohydrate Interaction Kinetics

Biotin-Streptavidin Enzyme-linked Immunosorbent Assay for the Determination of Dibutyl Phthalate in Beverages and Drinking Water Using a Specific Polyclonal Antibody

Enzyme-Linked Competitive Binding Assays Based on the Biotin/Avidin Interaction

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