Coordination-driven reversible surfaces with site-specifically immobilized nanobody for dynamic cancer cell capture and release
Coordination-driven reversible surfaces with site-specifically immobilized nanobody for dynamic cancer cell capture and release
复制标题
具有位点特异性固定纳米体的协调驱动的可逆表面，用于动态癌细胞捕获和释放
DOI：
10.1039/d0tb00574f
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发表时间：
2020
影响因子：
7
通讯作者：
Jia Lingyun
中科院分区：
文献类型：
--
作者：
Han Lulu;Peng Ruilian;Jiang Wenning;Xu Ting;Zhang Chong;Chen Kaidi;Zhang Yixin;Song Haiyuan;Jia Lingyun
从血液中选择性分离循环肿瘤细胞（CTC）为癌症患者的诊断、预后和个性化治疗提供了一种非侵入性途径。CTC的特异性捕获通常基于抗体与细胞膜上的受体之间的免疫亲和识别。然而，由于材料表面上的大抗体的有限负载能力，使用传统抗体高效分离CTC仍然是一个挑战。在本文中，使用小尺寸的纳米抗体（Nb），我们开发了一种广泛适用的策略来构建可逆的位点特异性固定的Nb表面，用于捕获和释放表皮样癌细胞系A431细胞。纳米抗体（Nb）的组氨酸标签（His-标签）与螯合至NTA修饰的聚（甲基丙烯酸2-羟乙酯）（PHEMA）刷的Ni 2+离子之间的配位相互作用用于实现EGFR Nb（PHEMA-aEGFR表面）的位点特异性固定。具有最大化活性的高密度固定化纳米抗体导致在仅30 min内高效捕获81%的稀有A431细胞，与固定在平坦表面上的常规抗体相比，显示出更高的捕获产率和更短的捕获时间。此外，PHEMA-aEGFR表面表现出低捕获极限（1个细胞mL-1）、对捕获细胞的细胞相容性以及可忽略的PBMC非特异性粘附。使用咪唑进行竞争配位的一步处理，86%的捕获细胞被有效释放。这种多功能和动态的位点特异性固定化纳米抗体策略为以低成本高效捕获和释放稀有细胞的材料和仪器的开发铺平了新的道路。
Selective isolation of circulating tumor cells (CTCs) from blood provides a non-invasive avenue for the diagnosis, prognosis and personalized treatment for patients with cancer. The specific capture of CTCs is conventionally based on the immunoaffinity recognition between antibody and receptor on cell membranes. However, using a traditional antibody for high-efficiency isolation of CTCs remains a challenge due to the limited loading capacity of the large antibodies on material surfaces. Herein, using a small-sized nanobody (Nb), we developed a widely applicable strategy to construct reversible site-specifically immobilized Nb surfaces for the capture and release of epidermoid cancer cell line A431 cells. Coordination interaction between the histidine tag (His-tag) of the nanobody (Nb) and Ni2+ ions that chelated to the NTA-modified poly(2-hydroxyethyl methacrylate) (PHEMA) brushes was used to achieve site-specific immobilization of EGFR Nb (PHEMA-aEGFR surfaces). The high-density immobilized nanobody possessing maximized activity resulted in the high-efficiency capture of 81% rare A431 cells within just 30 min, showing a higher capture yield and shorter capture time compared with that achieved by the conventional antibody immobilized on the flat surface. Additionally, the PHEMA-aEGFR surfaces exhibited low capture limit (1 cell mL−1), cytocompatibility for captured cells, as well as negligible non-specific adhesion of PBMCs. With a one-step treatment using imidazole for competitive coordination, 86% of the captured cells were effectively released. This multifunctional and dynamic site-specifically immobilized nanobody strategy paves a new path in the development of materials and instruments for the high-efficiency capture and release of rare cells at a low cost.