Immunoassay-type biosensor based on magnetic nanoparticle capture and the fluorescence signal formed by horseradish pero
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ORIGINAL PAPER
Immunoassay-type biosensor based on magnetic nanoparticle capture and the fluorescence signal formed by horseradish peroxidase catalysis for tumor-related exosome determination Hao Chen 1 & Dan Luo 1 & Bing Shang 1 & Jianjun Cao 1 & Jian Wei 1 & Qinhua Chen 1,2 & Jishun Chen 1 Received: 11 September 2019 / Accepted: 13 April 2020 # Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract A sandwich-type fluorescent biosensor for the determination of tumor-related exosome was designed. It is based on magnetic nanoparticle (MNP) capture and horseradish peroxidase (HRP) catalysis. MNPs were used as the substrate to capture exosomes by modifying the CD63 antibody on MNPs surface. After that, the biotinylated epithelial cell adhesion molecule (EpCAM) antibody was used to capture the tumor-related exosomes, which specifically express EpCAM. A novel method for the fluorescence measurement of tumor-associated exosome was achieved, with a detection limit as low as 200 (± 9) particles mL−1. The analytical range of this method is from 576 (± 15) particles mL−1 to 5.76 × 107 (± 5.1 × 105) particles mL−1. For the fluorescence measurement, the excitation wavelength was set to 320 nm. Fluorescent spectra were collected at emission wavelength in the range 370 to 550 nm; the data shown in the calibration plot were studied by using the fluorescence intensity at 406 nm. This sensor was also able to successfully detect the exosomes from the plasma of patients with hepatocellular carcinoma (HCC) and healthy humans. Keywords Fluorescent biosensor . Cancer diagnosis . Sandwich structure . Biomarker . CD63 . EpCAM
Introduction With the increase in the incidence of cancer, early detection is the most effective way to reduce cancer mortality in humans [1]. Tissue biopsy has always been the gold standard diagnostic procedure for tumors [2]. However, tissue biopsy has some
Hao Chen and Dan Luo contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00604-020-04275-x) contains supplementary material, which is available to authorized users. * Qinhua Chen [email protected] * Jishun Chen [email protected] 1
Affiliated Dongfeng Hospital, Hubei University of Medicine, Shiyan 442008, Hubei, China
2
Department of Pharmacy, Shenzhen Baoan Authentic TCM Therapy Hospital, Shenzhen 518101, Guangdong, China
limitations, such as their invasiveness and inability to reflect the spatial heterogeneity of tumors [3]. Liquid biopsy has gradually been considered as an approach for clinical diagnosis of cancers. It is a technique that utilizes circulating biomarkers in the body fluids of cancer patients to provide information about cancer [4]. It focuses on the detection of circulating tumor cells, circulating tumor DNA and exosomes [5]. Liquid biopsy can be used to analyze the molecular characterization of the tumor without the need for a tissue biopsy. It is vitally important to choose the specific biomarkers in liquid biopsies, and the exosome is one of the mos
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