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Concentration-Normalized Electroanalytical Assaying of Exosomal Markers.
Anal Chem. 2017 03 07; 89(5):3184-3190.AC

Abstract

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-to-noise. 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 × 105 particles mL-1 (equivalent to 320 aM or 9500 exosomes in 50 μL) for intact exosomes and 3-5 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.

Authors+Show Affiliations

Department of Chemistry, Physical and Theoretical Chemistry Laboratory, University of Oxford , South Parks Road, Oxford OX1 3QZ, United Kingdom.Nuffield Department of Clinical Neurosciences, University of Oxford , John Radcliffe Hospital, Oxford OX3 9DU, United Kingdom.Department of Chemistry, Physical and Theoretical Chemistry Laboratory, University of Oxford , South Parks Road, Oxford OX1 3QZ, United Kingdom.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

28192902

Citation

Li, Qian, et al. "Concentration-Normalized Electroanalytical Assaying of Exosomal Markers." Analytical Chemistry, vol. 89, no. 5, 2017, pp. 3184-3190.
Li Q, Tofaris GK, Davis JJ. Concentration-Normalized Electroanalytical Assaying of Exosomal Markers. Anal Chem. 2017;89(5):3184-3190.
Li, Q., Tofaris, G. K., & Davis, J. J. (2017). Concentration-Normalized Electroanalytical Assaying of Exosomal Markers. Analytical Chemistry, 89(5), 3184-3190. https://doi.org/10.1021/acs.analchem.6b05037
Li Q, Tofaris GK, Davis JJ. Concentration-Normalized Electroanalytical Assaying of Exosomal Markers. Anal Chem. 2017 03 7;89(5):3184-3190. PubMed PMID: 28192902.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Concentration-Normalized Electroanalytical Assaying of Exosomal Markers. AU - Li,Qian, AU - Tofaris,George K, AU - Davis,Jason J, Y1 - 2017/02/10/ PY - 2017/2/15/pubmed PY - 2019/1/3/medline PY - 2017/2/15/entrez SP - 3184 EP - 3190 JF - Analytical chemistry JO - Anal Chem VL - 89 IS - 5 N2 - 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-to-noise. 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 × 105 particles mL-1 (equivalent to 320 aM or 9500 exosomes in 50 μL) for intact exosomes and 3-5 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. SN - 1520-6882 UR - https://www.unboundmedicine.com/medline/citation/28192902/Concentration_Normalized_Electroanalytical_Assaying_of_Exosomal_Markers_ L2 - https://doi.org/10.1021/acs.analchem.6b05037 DB - PRIME DP - Unbound Medicine ER -