Folic acid-functionalized magnetic nanoprobes via a PAMAM dendrimer/SA-biotin mediated cascade-amplifying system for the efficient enrichment of circulating tumor cells
Folic acid-functionalized magnetic nanoprobes via a PAMAM dendrimer/SA-biotin mediated cascade-amplifying system for the efficient enrichment of circulating tumor cells
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通过 PAMAM 树枝状聚合物/SA-生物素介导的级联放大系统的叶酸功能化磁性纳米探针可有效富集循环肿瘤细胞
DOI：
10.1039/d0bm01212b
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发表时间：
2020
影响因子：
6.6
通讯作者：
Wang Zhifei
中科院分区：
文献类型：
--
作者：
Meng Xiangyu;Sun Pingfeng;Xu Hengyi;Wang Zhifei
循环肿瘤细胞（CTC）作为一种液体活检物，对于肿瘤的早期诊断、及时治疗和转移复发的实际评估具有重要意义。然而，在复杂血液样品中富集稀有CTC仍然是一个重大挑战。在此，构建了独特且高度灵敏的叶酸（FA）官能化级联扩增系统修饰的磁性纳米颗粒（MNP），以有效地捕获全血中的CTC。在该系统中，作为靶向分子，许多FA分子通过聚乙二醇（PEG）接头缀合在PAMAM树枝状聚合物（PAMAM-FA）（第一扩增）的表面上，这可以促进卵巢癌细胞（SKOV 3细胞）表面上的叶酸受体（FR）的更容易结合。在此基础上，利用生物素对PAMAM-FA进行修饰，制备了生物素-PAMAM-FA（BPF）复合物，该复合物可与链霉亲和素（SA）修饰的MNP（SMs）通过SA-生物素系统进行联合收割机结合，从而有效地靶向和分离CTCs。所构建的MNPs-SA生物素-PAMAM-FA（SM@BPF）纳米探针的捕获效率为90.3%，具有高的细胞存活率（约93.2%）和最小的非特异性吸附（约25%）。此外，与SM/BPF捕获的SKOV 3细胞（两步法）相比，SM@ BPF捕获的SKOV 3细胞（一步法）表面吸附的纳米探针较少，这有利于进一步的生物学分析。我们期望这种基于识别分子的级联扩增系统将为卵巢癌的早期诊断提供创新的CTC富集平台。
As a liquid biopsy, circulating tumor cells (CTCs) have great significance for the early diagnosis, timely treatment, and practical evaluation of metastasis or recurrence of cancer. However, the enrichment of rare CTCs in complex blood samples is still a significant challenge. Here, unique and highly sensitive folic acid (FA)-functionalized cascade amplification system-modified magnetic nanoparticles (MNPs) were constructed to effectively capture CTCs in whole blood. In this system, as a targeted molecule, numerous FA molecules were conjugated on the surface of PAMAM dendrimers (PAMAM-FA) (first amplification) through a polyethylene glycol (PEG) linker, which could promote the more facile binding of folate receptors (FR) on the surface of ovarian cancer cells (SKOV3 cells). Then, PAMAM-FA was further modified with biotin to fabricate biotin-PAMAM-FA (BPF), which could combine with streptavidin (SA)-modified MNPs (SMs) via the SA-biotin system to efficiently target and separate CTCs. The capture efficiency of the constructed MNPs-SA ∼ biotin-PAMAM-FA (SM@BPF) nanoprobes was 90.3% with high cell viability (∼93.2%) and minimal non-specific adsorption (∼25%). Moreover, fewer nanoprobes were absorbed on the surface of the SM@BPF-captured SKOV3 cells (one-step method) compared with the SM/BPF-captured SKOV3 cells (two-step method), which was beneficial for further biological analysis. We expect that this recognition molecule-based cascade amplification system will provide an innovative CTCs enrichment platform for the early-stage diagnosis of ovarian cancer.