Biomarker Branch, National Cancer Center, 323 Ilsan-ro, Ilsan-dong-gu, Goyang, Gyeonggi 10408, Korea.
Department of Medical Science, Yonsei University College of Medicine, 50 Yonsei-Ro, Seodaemun-Gu, Seoul 03722, South Korea.
Theranostics. 2018 Jan 1;8(2):505-517. doi: 10.7150/thno.21967. eCollection 2018.
Recent developments in genomic and molecular methods have revolutionized the range of utilities of tumor-associated circulating biomarkers in both basic and clinical research. Herein, we present a novel approach for ultrasensitive extraction of cfDNA and CTCs, at high yield and purity, via the formation of magnetic nanowire networks. We fabricated and characterized biotinylated cationic polyethylenimine and biotinylated antibody cocktail-conjugated magnetic polypyrrole NWs (PEI/mPpy NW and Ab cocktail/mPpy NW, respectively). We applied these NWs to the extraction of cfDNA and CTC from the blood of 14 patients with lung cancer. We demonstrated reliable detection of mutations based on digital droplet PCR analysis of cfDNA and CTC DNA from patients with lung cancer. The NW networks confined with a high density of magnetic nanoparticles exhibited superior saturation magnetization, which enabled rapid and high-yield capture whilst avoiding or minimizing damage and loss. The NW networks enabled the co-isolation of CTCs and cfDNA of high quality and sufficient quantities, thus allowing the amplification of rare and low-prevalence cancer-related mutations. The simple, versatile, and highly efficient nanowire network tool allows sensitive and robust assessment of clinical samples.
近年来,基因组学和分子方法的发展彻底改变了肿瘤相关循环生物标志物在基础和临床研究中的应用范围。在此,我们提出了一种通过形成磁性纳米线网络来实现 cfDNA 和 CTC 高产量和高纯度超灵敏提取的新方法。我们制备并表征了生物素化阳离子聚乙烯亚胺和生物素化抗体鸡尾酒偶联的磁性聚吡咯纳米线(PEI/mPpy NW 和 Ab 鸡尾酒/mPpy NW)。我们将这些 NW 应用于从 14 名肺癌患者的血液中提取 cfDNA 和 CTC。我们通过对来自肺癌患者的 cfDNA 和 CTC DNA 的数字液滴 PCR 分析,证明了基于突变的可靠检测。具有高密度磁性纳米颗粒的 NW 网络表现出优越的饱和磁化强度,能够实现快速和高产率的捕获,同时避免或最小化损伤和损失。NW 网络能够共分离高质量和足够数量的 CTC 和 cfDNA,从而允许扩增罕见和低流行度的癌症相关突变。这种简单、多功能且高效的纳米线网络工具允许对临床样本进行敏感和稳健的评估。