Latex-based thin-layer immunoaffinity chromatography for quantitation of protein analytes

A rapid immunochromatographic method for qualitativeand quantitative analysis of protein antigens is described. The method is based on the ‘sandwich” assay format using monoclonal antibodies (Mabs) of two distinct specificities. Mabs of one specificity are covalently immobilized to a defined detecti...

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Veröffentlicht in:Analytical biochemistry 1992-10, Vol.206 (1), p.168-171
Hauptverfasser: Birnbaum, Staffan, Udén{xa, Catharina, Magnusson, Carl G.M., Nilsson, Staffan
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container_end_page 171
container_issue 1
container_start_page 168
container_title Analytical biochemistry
container_volume 206
creator Birnbaum, Staffan
Udén{xa, Catharina
Magnusson, Carl G.M.
Nilsson, Staffan
description A rapid immunochromatographic method for qualitativeand quantitative analysis of protein antigens is described. The method is based on the ‘sandwich” assay format using monoclonal antibodies (Mabs) of two distinct specificities. Mabs of one specificity are covalently immobilized to a defined detection zone on a porous membrane while Mabs of the other specificity are covalently coupled to blue latex particles which serve as a label. The sample is mixed with the Mab-coated particles and allowed to react. The mixture is then passed along a porous membrane by capillary action past the Mabs in the detection zone, which will bind the particles which have antigen bound to their surface, giving a blue color within this detection zone with an intensity logarithmetrically proportional to the antigen concentration in the sample. Analysis is complete in less than 10 min, requires a minimum amount of sample (4 μl), and has a detection limit below the nanomolar range for the antigen we studied, human chorionic gonadotropin.
doi_str_mv 10.1016/S0003-2697(05)80028-4
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subjects affinity chromatography
Analytical, structural and metabolic biochemistry
Antibody Specificity
Biological and medical sciences
chorionic gonadotropin
Chromatography, Affinity - methods
Chromatography, Thin Layer - methods
Fundamental and applied biological sciences. Psychology
Haptens - analysis
Humans
Immunoassay
Latex
man
Proteins
Proteins - analysis
quantitation
Sensitivity and Specificity
title Latex-based thin-layer immunoaffinity chromatography for quantitation of protein analytes
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