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Title 

Human alpha-fetal protein immunoassay using fluorescence suppression with fluorescent-bead/antibody conjugate and enzymatic reaction

Authors 

J AhnYong Beom ShinJ LeeM G Kim

Publisher 

Elsevier

Issue Date 

2015

Citation 

Biosensors & Bioelectronics, vol. 71, no. 0, pp. 115-120

Keywords 

Alpha-fetal protein (AFP)Enzyme-catalyzed precipitationFluorescence suppressionFluorescent immunoassay

Abstract 

The aim of the study was to develop a simple and rapid immunoassay using fluorescent microbeads and enzyme-substrate reactions to measure alpha-fetal protein (AFP) concentrations. We demonstrated the functionality of the fluorescent immunosensor using antibody-conjugated fluorescent latex beads (AB-FLBs) and horseradish peroxidase (HRP) to catalyze a reaction, where the products would precipitate and suppress the fluorescence of AB-FLBs. First, the AB-FLBs were incubated with antigen, biotinylated antibodies (bABs), and streptavidin-HRP (SAv-HRP) to form a sandwich-type immunoreaction. The mixture was then filtered through a membrane to concentrate the beads on a small area. After washing to remove unbound bABs and SAv-HRP, a chromogenic HRP substrate and H2O2 were added to form precipitates on the FLB surface. The suppression of the fluorescence was measured with a fluorescent image analyzer system. Under optimized conditions, AFP could be measured at concentrations as low as 1 pg mL-1 with a dynamic range up to 100 ng mL-1

ISSN 

0956-5663

Link 

http://dx.doi.org/10.1016/j.bios.2015.03.073

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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