文献类型：期刊文献

英文题名：Direct Exosome Quantification via Bivalent-Cholesterol-Labeled DNA Anchor for Signal Amplification

作者：He, Fang[1];Liu, Hui[2];Guo, Xinggang[2];Yin, Bin-Cheng[1];Ye, Bang-Ce[1,3]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Lab Biosyst & Microanal, Shanghai 200237, Peoples R China;[2]Eastern Hepatobiliary Surg Hosp, Dept Hepat Surg 3, Shanghai 200438, Peoples R China;[3]Zhejiang Univ Technol, Coll Pharmaceut Sci, Collaborat Innovat Ctr Yangtze River Delta Reg Gr, Hangzhou 310014, Zhejiang, Peoples R China

年份：2017

卷号：89

期号：23

起止页码：12968

外文期刊名：ANALYTICAL CHEMISTRY

收录：;EI(收录号：20175004529339);WOS:【SCI-EXPANDED(收录号:WOS:000417549600051)】；

基金：This work was jointly supported by the National Natural Science Foundation of China (Grants 21335003, 21675052, 21575089, and 81271649), the Fundamental Research Funds for the Central Universities, the Science Fund for Creative Research Groups (Grant 21421004), and Programme of Introducing Talents of Discipline to Universities (Grant B16017).

语种：英文

外文关键词：Bioassay - Antigen-antibody reactions - Signal processing

摘要：Exosomes, as an important subpopulation of extracellular vesicles (EVs), play an important role in intercellular communications in various important pathophysiological processes, especially cancer-related. However, reliable and convenient quantitative methods for their determination are still technically challenging. In this study, we developed an efficient and direct method by combining immunoaffinity and lipid membrane surface modification into a single platform for specific isolation and accurate quantification of exosomes. Exosomes are specifically captured by immunomagnetic beads, and then a bivalent-cholesterol (BChol)-labeled DNA anchor with high affinity is spontaneously inserted into the exosome membrane. The rationally designed sticky end of the anchor acts as the initiator for the subsequent horseradish peroxidase (HRP)-linked hybridization chain reaction (HCR) for signal amplification. Detection is based on the color change of HRP-catalyzed H2O2,-mediated oxidation of 3,3',5,5'- tetramethyl benzidine (TMB), which can be conveniently observed by the naked eye and monitored by UV vis spectrometry. This proposed method enables sensitive detection of 2.2 X 103 exosomes per microliter with a relative standard deviation of <5.6%, with 100-fold higher sensitivity compared to conventional ELISA. We believe that our assay has considerable potential as a routine bioassay (cost-efficient, reliable, and easy to operate) for the accurate quantification of exosomes in clinical samples.