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使用线粒体DNA包裹的单壁碳纳米管比色法检测免疫磁捕获的少量循环肿瘤细胞

Colorimetric detection of immunomagnetically captured rare number CTCs using mDNA-wrapped single-walled carbon nanotubes.

作者信息

Zhu Liang, Feng Xingqing, Yang Shenhao, Wang Jianyun, Pan Yixin, Ding Jinhua, Li Chenglin, Yin Xiaoxing, Yu Yanyan

机构信息

Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, 209 Tongshan Road, Xuzhou, 221004, Jiangsu, PR China.

Department of Neurosurgery, Center for Functional Neurosurgery Ruijin Hospital, Shanghai Jiaotong Univeristy School of Medicine, 197 Ruijin Second Road, Huangpu, Shanghai, 200025, PR China.

出版信息

Biosens Bioelectron. 2021 Jan 15;172:112780. doi: 10.1016/j.bios.2020.112780. Epub 2020 Oct 29.

Abstract

The number of CTCs in peripheral blood is of great significance for the early diagnosis, recurrence and prognosis evaluation of tumor patients. Consequently, it is required to develop simple and effective technique to realize the capture and detection of rare number CTCs. Herein, a SiO, gelatin and biotinylated EpCAM aptamer P1 modified FeO immunomagnetic nanoparticles (IMNs) were prepared for the specific capture and nondestructive release of trace amounts of CTCs. Then, utilizing the peroxidase-like activity of single-walled carbon nanotubes (SWCNTs) and the effect of non-specific DNA sequences on this activity, a colorimetric probe was constructed by modifying the three DNA sequences (mDNA) onto the IMNs. When target cell was present, due to the specific interaction between cells and P1, the conformational structure of P1 was changed. Consequently, the mDNA linked with P1 on IMNs was replaced by the cell and released from IMNs. In this way, the quantification of captured CTCs could be converted to that of released mDNA. This strategy combined the capture and detection of CTCs as a whole and could detect down to 10 cells with a high selectivity. Finally, we achieved the accurate quantification of CTCs in lysed bloods from 12 clinical tumor patients, which exhibited a great promise for further clinical applications.

摘要

外周血中循环肿瘤细胞(CTCs)的数量对肿瘤患者的早期诊断、复发及预后评估具有重要意义。因此,需要开发简单有效的技术来实现对少量CTCs的捕获与检测。在此,制备了一种二氧化硅、明胶和生物素化上皮细胞黏附分子适配体P1修饰的四氧化三铁免疫磁性纳米颗粒(IMNs),用于特异性捕获和无损释放痕量CTCs。然后,利用单壁碳纳米管(SWCNTs)的过氧化物酶样活性以及非特异性DNA序列对该活性的影响,通过在IMNs上修饰三条DNA序列(mDNA)构建了一种比色探针。当存在靶细胞时,由于细胞与P1之间的特异性相互作用,P1的构象结构发生改变。因此,与IMNs上P1相连的mDNA被细胞取代并从IMNs上释放出来。通过这种方式,捕获的CTCs的定量可转化为释放的mDNA的定量。该策略将CTCs的捕获与检测整合为一体,能够以高选择性检测低至10个细胞。最后,我们实现了对12例临床肿瘤患者裂解血中CTCs的准确定量,这为进一步的临床应用展现出巨大潜力。

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