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MiRNA Extraction from Cell-Free Biofluid Using Protein Corona Formed around Carboxyl Magnetic Nanoparticles
ACS Biomaterials Science & Engineering ( IF 5.4 ) Pub Date : 2018-01-03 00:00:00 , DOI: 10.1021/acsbiomaterials.7b00668 Shengqiang Xu 1 , Seyedmehdi Hossaini Nasr 2 , Daoyang Chen 2 , Xiaoxian Zhang 3 , Liangliang Sun 2 , Xuefei Huang 2 , Chunqi Qian 1
ACS Biomaterials Science & Engineering ( IF 5.4 ) Pub Date : 2018-01-03 00:00:00 , DOI: 10.1021/acsbiomaterials.7b00668 Shengqiang Xu 1 , Seyedmehdi Hossaini Nasr 2 , Daoyang Chen 2 , Xiaoxian Zhang 3 , Liangliang Sun 2 , Xuefei Huang 2 , Chunqi Qian 1
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MicroRNA (miRNA) in urine has been considered as a potential biomarker for early stage diagnosis of multiple diseases like urinary system cancer, kidney injury, and diabetes because of their many demonstrated advantages including long-term stability and noninvasiveness. However, the traditional enrichment and extraction processes of miRNAs from urine are cumbersome and tedious due to the low concentration and multiple carriers of miRNAs. Herein, we present a novel method to collect low concentrations of miRNAs from dilute solutions such as urine and cell culture medium. Ten nanometer core sized magnetic nanoparticles with carboxylic acid coating can adsorb low-concentration proteins, and form protein corona which makes them easy to aggregate and precipitate for subsequent isolation. In urine and cell culture medium, these nanoparticles can aggregate with proteins, including miRNAs-associated protein Argonaute 2 and microvesicle-related proteins, to form precipitates, so that miRNAs can be easily extracted from pellets by small amount of lysis buffer for subsequent analysis such as real-time PCR. Our method provides a facile way to enrich miRNAs from biofluids without the need of ultracentrifugation and immunoprecipitations, bringing remarkable convenience to miRNA-based biomarker research.
中文翻译：
使用围绕羧基磁性纳米粒子形成的蛋白质电晕，从无细胞生物流体中提取MiRNA。
尿液中的MicroRNA（miRNA）被认为是多种疾病（如泌尿系统癌症，肾脏损伤和糖尿病）早期诊断的潜在生物标志物，因为它们具有许多优点，包括长期稳定性和无创性。然而，由于miRNA的低浓度和多种载体，传统的从尿液中富集和提取miRNA的过程繁琐且乏味。在本文中，我们提出了一种从稀溶液（如尿液和细胞培养基）中收集低浓度miRNA的新颖方法。十个具有羧酸涂层的核纳米大小的磁性纳米粒子可以吸附低浓度的蛋白质，并形成蛋白质电晕，使它们易于聚集和沉淀，以用于后续分离。在尿液和细胞培养基中，这些纳米颗粒可以与蛋白质聚集，包括与miRNA相关的蛋白质Argonaute 2和与微泡相关的蛋白质，从而形成沉淀物，从而可以通过少量裂解缓冲液轻松地从沉淀物中提取miRNA，以进行后续分析（例如实时PCR）。我们的方法提供了一种从生物流体中富集miRNA的简便方法，而无需超速离心和免疫沉淀，这为基于miRNA的生物标志物研究带来了极大的便利。
更新日期：2018-01-03
中文翻译：
使用围绕羧基磁性纳米粒子形成的蛋白质电晕，从无细胞生物流体中提取MiRNA。
尿液中的MicroRNA（miRNA）被认为是多种疾病（如泌尿系统癌症，肾脏损伤和糖尿病）早期诊断的潜在生物标志物，因为它们具有许多优点，包括长期稳定性和无创性。然而，由于miRNA的低浓度和多种载体，传统的从尿液中富集和提取miRNA的过程繁琐且乏味。在本文中，我们提出了一种从稀溶液（如尿液和细胞培养基）中收集低浓度miRNA的新颖方法。十个具有羧酸涂层的核纳米大小的磁性纳米粒子可以吸附低浓度的蛋白质，并形成蛋白质电晕，使它们易于聚集和沉淀，以用于后续分离。在尿液和细胞培养基中，这些纳米颗粒可以与蛋白质聚集，包括与miRNA相关的蛋白质Argonaute 2和与微泡相关的蛋白质，从而形成沉淀物，从而可以通过少量裂解缓冲液轻松地从沉淀物中提取miRNA，以进行后续分析（例如实时PCR）。我们的方法提供了一种从生物流体中富集miRNA的简便方法，而无需超速离心和免疫沉淀，这为基于miRNA的生物标志物研究带来了极大的便利。
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