Journal
ACS SENSORS
Volume 4, Issue 1, Pages 93-99Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssensors.8b00949
Keywords
ELISA; ultrasensitive assay; expression immunoassay; cell-free protein synthesis; E. coli extract; AFP
Funding
- National Research Foundation of Korea [2016M1A5A1027465, 2014M3C1A3051473]
- BioNano Health Guard Research Center - Ministry of Science and ICT (MSIT) of Korea as a Global Frontier Project [H-GUARD_2013M3A6B2078950]
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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 a-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.
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