Detection System for Membrane Immunoassay Based on the Trapping of a Highly Colored Intermediate of the Peroxidase Reaction

Yatsimirskaya, Eugenia A.; Gavrilova, E. M.; Egorov, A. M.

doi:10.1006/abio.1993.1269

Cited by 5 publications

(2 citation statements)

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“…Colorimetric detection was based on the stabilisation of a highly coloured intermediate of OD oxidation in the presence of dextran sulfate [10]; its advantages for the membrane tests have been established previously [8]. Green spots (intermediate) were observed for 4-n-nonylphenol concentrations from 0 to 10 ng mL -1 whereas concentrations from 100 ng mL -1 resulted in a brown colour (final product) for the spot (data not presented).…”

Section: Membrane Assay Of 4-n-nonylphenolmentioning

confidence: 98%

Determination of 4- n -nonylphenol in water by enzyme immunoassay

2003

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A range of monoclonal antibody-based competitive immunoassays in the format of microtitre plate ELISA and dipstick tests for quantitative and semi-quantitative detection of 4- n -nonylphenol in water was developed. A simple visual dipstick test was based on changing of spot colour from green to brown in the presence of 4- n -nonylphenol at concentrations within the range 10-100 ng mL(-1). Two different detection systems were used for quantitative immunoassay. Application of enhanced chemiluminescence (ECL) resulted in an increase of the sensitivity of ELISA when compared to conventional colorimetric detection. Thus a detection limit of 0.06 ng mL(-1 )of 4- n -nonylphenol was achieved with IC(50) 2.0 ng mL(-1). The tests developed were applied to natural and spiked water samples.

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Section: Membrane Assay Of 4-n-nonylphenolmentioning

confidence: 98%

Determination of 4- n -nonylphenol in water by enzyme immunoassay

2003

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“…The first system, including TMB as a chromogen and dioctylsulphosuccinate as a precipitating agent, was recently reported to be one of the most sensitive for a dipstick assay (Giersch, 1993). The second system was based on the stabilization of a highly coloured intermediate of OD oxidation in the presence of dextrane sulphate (Yatsimirskaya et al, 1993). Its advantages for the membrane immunoassay tests had been established previously (Rubtsova & Gavrilova, 1994).…”

Section: Membrane Strip Assay For 24-dmentioning

confidence: 99%

Chemiluminescent immunoassay: Application of a portable scanning luminometer for the determination of 2,4‐dichlorophenoxyacetic acid in microtiter and membrane strip format

Rubtsova

Wittmann

Egorov

et al. 1997

Food and Agricultural Immunology

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A sensitive competitive immunoassay with chemiluminescent detection for the determination of 2,4-dichlorophenoxyacetic acid (2,4-D) was carried out in a microtiter and membrane strip format. The application of a portable scanning luminometer for the detection of light emission in polystyrene strips and from the surface of a porous membrane resulted in an increase in the sensitivity of the assay and a wider detection range when compared to conventional colorimetric detection. With the assay in microtiter strip format, 2,4-D can be determined within the range 0.005-1 μg l -1 with an IC 50 at 0.04 p.g l -1 . Different immobilization techniques involving adsorption and covalent immobilization on different kinds of modified nylon were examined to optimize the attachment of specific antibodies to the surface of porous membranes. Photoimmobilization on positively charged nylon modified with p-azidobenzoic acid was found to result in higher sensitivity. The application of a membrane strip format allows the quick quantitative detection of 2,4-D at concentrations above 0.02 μg l -1 . Both tests in microtiter and membrane strip formats can be performed under field conditions for the screening of water samples.

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Investigation of several parameters influencing signal generation in flow-through membrane-based enzyme immunoassay

et al. 2006

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Rapid-response analytical tests that can be performed at the point of sampling are based on a visual detection system. The influence of different factors on the signal generation in a membrane-based enzyme immunoassay was investigated. The research was applied to a flow-through immunoassay for the detection of ochratoxin A (OTA). This assay format is a very convenient, simple and fast qualitative screening tool. Conjugates of OTA with horseradish peroxidase (HRP) and alkaline phosphatase (AP) were used as enzyme tracers. A new conjugate OTA-AP has been synthesized in our laboratory and its performance in the assay was compared with that of OTA-HRP. Different substrate systems for HRP and AP were compared. Several reagents, including polymers and surfactants, were tested for their possible effect on signal generation with the use of OTA-HRP conjugate. Polymers such as poly(vinyl alcohol) (PVA) and poly(ethylene glycol) (PEG) 6000 exerted a favourable effect on signal amplification, whereas surfactants negatively affected assay performance. The highest signal amplification (30-70% compared to the standard assay procedure) was achieved using 0.5% PVA in tetramethylbenzidine (TMB) Colorburst substrate solution and phosphate-buffered saline (PBS) for the washing step. It allowed more reliable visual estimation of the results from OTA-HRP assay. Exclusion of the detergent (Tween 20) from the washing solution exerted a favourable effect on assay performance using both enzyme tracers. The assay using OTA-HRP was more susceptible to matrix interferences than the assay with OTA-AP. Signal development in the matrix was better for the OTA-AP assay and visual estimation of the results was easier to perform in this case. For the analysis of spiked wheat samples, OTA-AP conjugate gave a more sensitive, stable and reproducible assay with a cut-off level of 4 microg kg(-1) for OTA. The application of the new OTA-AP conjugate resulted in improved assay performance for the food samples.

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Detection System for Membrane Immunoassay Based on the Trapping of a Highly Colored Intermediate of the Peroxidase Reaction

Cited by 5 publications

References 0 publications

Determination of 4- n -nonylphenol in water by enzyme immunoassay

Determination of 4- n -nonylphenol in water by enzyme immunoassay

Chemiluminescent immunoassay: Application of a portable scanning luminometer for the determination of 2,4‐dichlorophenoxyacetic acid in microtiter and membrane strip format

Investigation of several parameters influencing signal generation in flow-through membrane-based enzyme immunoassay

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