期刊
ANALYTICAL CHEMISTRY
卷 80, 期 22, 页码 8477-8484出版社
AMER CHEMICAL SOC
DOI: 10.1021/ac801211s
关键词
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资金
- National Institutes of Health Counter ACT Program through the National Institute of Neurological Disorders and Stroke [U01 NS058161]
- CDC/NIOSH [R01 OH008173]
- DOE [DE-AC05-76RL01830]
A new magnetic electrochemical immunoassay has been developed as a tool for biomonitoring exposures to organophosphate (OP) compounds, e.g., insecticides and chemical nerve agents, by directly detecting organophosphorylated acetylcholinesterase (OP-AChE). This immunoassay uniquely incorporates highly efficient magnetic separation with ultrasensitive square wave voltammetry (SWT) analysis with quantum dots (QDs) as labels. A pair of antibodies was used to achieve the specific recognition of OP-AChE that was prepared with paraoxon as an OP model agent. Antiphosphoserine polyclonal antibodies were anchored on amorphous magnetic particles preferably chosen to capture OP-AChE from the sample matrixes by binding their phosphoserine moieties that were exposed through unfolding the protein adducts. Ibis was validated by electrochemical examinations and enzyme-linked inummosorbent assays. Furthermore, antihuman AChE monoclonal antibodies were labeled with cadmium-source QDsto selectively recognize the captured OP-AChE, as characterized by transmission electron microscopy. The subsequent electrochemical SWV analysis of the cadmium component released by acid from the coupled QDs was conducted on disposable screen-printed electrodes. Experimental results indicated that the SWV-based immunoassays could yield a linear response over a broad concentration range of 0.3-300 ng/mL OP-AChE in human plasma with a detection limit of 0.15 ng/mL Such a novel electrochemical immunoassay holds great promise as a simple, selective, sensitive, and field-deployable tool for the effective biomonitoring and diagnosis of potential exposures to nerve agents and pesticides.
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