背景:捕获循环肿瘤细胞(CTC)的技术在诊断中起着至关重要的作用,疗效评估,并对肺癌的预后进行预测。然而,复杂血液环境的存在通常会导致血细胞严重的非特异性蛋白质吸附和干扰,这对CTC捕获的特异性产生负面影响。非常需要开发用于CTC捕获的新型纳米材料,其具有来自蛋白质或血细胞的显著的抗非特异性吸附。
结果:我们提出了一种新型的免疫磁性探针Fe3O4@(PEI/AA)4@Apt。通过逐层组装,用四层PEI/AA复合材料对Fe3O4颗粒表面进行改性。此外,在Fe3O4@(PEI/AA)4上同时连接靶向上皮标记物EpCAM(SYL3C)和间充质标记物CSV(ZY5C)的适体,以提高对不同表型CTC的检测并减少假阴性。结果表明,(PEI/AA)4涂层不仅使非特异性蛋白质吸附最小化,而且还将红细胞的吸附率显着降低到仅1%,因此,Fe3O4@(PEI/AA)4@Apt探针对CTC的捕获效率非常高(95.9%)。在随后的临床样本验证中,该探针还可有效捕获肺癌患者的罕见CTC.
■通过逐层自组装技术制备了具有可控层的(PEI/AA)聚合复合材料,对蛋白质和细胞表现出显著的抗非特异性吸附能力。重要的是,Fe3O4@(PEI/AA)4@Apt探针可显著提高肺癌患者CTCs捕获纯度,达到89.36%。第一次,这项研究结合了可控(PEI/AA)层与磁分离,以创新地建立一个电阻界面,显着提高CTC的特定捕获性能,拓宽这种聚合复合材料的应用。
BACKGROUND: The technology of capturing circulating tumor cells (CTCs) plays a crucial role in the diagnosis, evaluation of therapeutic efficacy, and prediction of prognosis in lung cancer. However, the presence of complex blood environment often results in severe nonspecific protein adsorption and interferences from blood cells, which negatively impacts the specificity of CTCs capture. There is a great need for development of novel nanomaterials for CTCs capture with prominent anti-nonspecific adsorptions from proteins or blood cells.
RESULTS: We present a novel immune magnetic probe Fe3O4@(PEI/AA)4@Apt. The surface of Fe3O4 particles was modified with four layers of PEI/AA composite by layer-by-layer assembly. Furthermore, aptamers targeting epithelial marker EpCAM (SYL3C) and mesenchymal marker CSV (ZY5C) were simultaneously connected on Fe3O4@(PEI/AA)4 to improve the detection of different phenotypic CTCs and reduce false negatives. The results demonstrated that the (PEI/AA)4 coatings not only minimized non-specific protein adsorptions, but also significantly reduced the adsorption rate of red blood cells to a mere 1 %, as a result of which, the Fe3O4@(PEI/AA)4@Apt probe achieved a remarkably high capture efficiency toward CTCs (95.9 %). In the subsequent validation of clinical samples, the probe was also effective in capturing rare CTCs from lung cancer patients.
UNASSIGNED: A (PEI/AA) polymerized composite with controllable layers was fabricated by layer-by-layer self-assembly technique, which displayed remarkable anti-nonspecific adsorption capabilities toward proteins and cells. Importantly, Fe3O4@(PEI/AA)4@Apt probe significantly improved CTCs capture purity in lung cancer patients to 89.36 %. For the first time, this study combined controllable (PEI/AA) layers with magnetic separation to innovatively build a resistant interface that significantly improves the specific capture performances of CTCs, broadening the application of this polymerized composite.