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Title 

Cascading amplification of immunoassay signal by cell-free expression of firefly luciferase from detection antibody-conjugated DNA in an Escherichia coli extract

Authors 

Ju Young ByunK H LeeYong Beom ShinD M Kim

Publisher 

American Chemical Society

Issue Date 

2019

Citation 

ACS Sensors

Keywords 

AFPE. coli extractELISAcell-free protein synthesisexpression immunoassayultrasensitive assay

Abstract 

An expression immunoassay is a powerful technique that combines unique features of immunosorbent assays and cell-free protein synthesis. The main advantage of the expression immunoassay is a greatly amplified signal, whereas a conventional enzyme-linked immunosorbent assay (ELISA) employs a single enzyme molecule conjugated to a detection antibody to produce a measurable signal. Expression immunoassays utilize a DNA molecule conjugated to a target-bound antibody to generate multiple enzyme molecules that then produce the signal. To date, expression immunoassays have not been widely adopted due to the limited availability of efficient methods for translating antibody-conjugated DNA. We developed a highly efficient translation module for expression immunoassays using an Escherichia coli extract-based cell-free protein synthesis system. When we used our immunoassay technique to detect α-fetoprotein, we achieved a limit of detection of 7 fM. Given the outstanding sensitivity that can be obtained with only minimal modifications to the procedure of standard ELISA, we believe that this method will open up new possibilities for widespread application of expression immunoassays to ultrasensitive detection and diagnostics.

URI 

https://doi.org/10.1021/acssensors.8b00949

ISSN 

2379-3694

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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