期刊
ANALYTICAL CHEMISTRY
卷 89, 期 5, 页码 3184-3190出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.6b05037
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资金
- EPSRC [EP/M006204/1]
- Wellcome Trust [097479/Z/11/Z]
- Oxford Biomedical Research Centre
- Alzheimers Research UK [ARUK-PPG2015B-2] Funding Source: researchfish
- Engineering and Physical Sciences Research Council [EP/M006204/1] Funding Source: researchfish
- Parkinson's UK [J-1403] Funding Source: researchfish
- EPSRC [EP/M006204/1] Funding Source: UKRI
- Wellcome Trust [097479/Z/11/Z] Funding Source: Wellcome Trust
Exosomes are both active in mediating intracellular communication and potentially present a potent cargo of disease biomarkers to an assay. The robust evaluation of exosomal markers could lead to a paradigm shift in clinical analysis and associated care. To date, much of this has been hindered by issues of sample preparation and assay signal-tonoise. We introduce here the use of ultrasensitive electrochemical impedance spectroscopy to quantify both external (tetraspanin) and internal (syntenin) exosome-specific markers. Associated exosome detection limits are 1.9 X 105 particles mL(-1) (equivalent to 320 aM or 9500 exosomes in 50 mu L) for intact exosomes and 3 S picomolar for internal exosomal syntenin levels with almost 5 decades of linear dynamic range. Sample preparation can be carried out by simple fine filtering of cell-conditioned medium prior to a non-NTA-determined (i.e., nanoparticle tracking analysis) exosome concentration analysis, lysing, and subsequent internal syntenin quantification. Such concentration-normalized dual-marker analysis can be used to define analytical zones in a manner which is then independent of absolute exosome concentration and sample preparation.
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