Development of Capture and CollectionMethods of Circulating Tumor Cells Utilizing Photocleavable Linkers

循环肿瘤细胞（CTCs）被定义为在外周血中发现的实体瘤起源的癌细胞。一般认为，这些ctc从患者的原发肿瘤上脱落，进入血液，在体内四处移动，然后附着在远处的组织上。因此，一般认为CTC的定量检测和分析对癌症患者的个人诊断是有用的。在本工作中，开发了一种捕获和回收活靶细胞的装置，该平台是用抗HEL抗体（抗HEL- igg， HEL=鸡蛋溶菌酶）通过光可切割的3-氨基-3-（2-硝基苯基）丙酸（ANP）连接剂修饰的硅（Si）晶片。利用衰减全反射傅里叶变换红外光谱（FTIR-ATR）和快速扫描原子力显微镜（FS-AFM）监测了硅片表面的修饰过程。通过FS-AFM和酶联免疫吸附试验（ELISA）观察了IgG在硅表面的附着及其通过ANP连接物光化学裂解的释放反应。此外，可以在硅片器件上选择性地收集在其细胞膜上表达HEL的SP2/O细胞（SP2/O-HEL）。ANP连接体的光化学裂解促进了活SP2/O细胞的有效释放，其活力通过三泛蓝染色实验得到验证。此外，还可以对恢复的细胞进行再培养。该方法是分离完整靶细胞的有效方法，适用于生物医学及相关领域。

Circulating tumor cells (CTCs) have been defined as cancer cells of solid tumor origin found in the peripheral blood. It isgenerally considered that these CTCs detach from primary tumors of patients, go into the bloodstream, travel around in the body, and then attach to the remote tissues. Accordingly, it is generally described that the quantitative detection and analysis of CTC is considered to be useful for the personal diagnosis of cancer patients. In this work, a device for the capture and recollection of live target cells is developed The platform was a silicon (Si) wafer modified with an anti-HEL antibody (anti-HEL-IgG, HEL= hen egg lysozyme) through a photocleavable 3-amino-3-(2-nitrophenyl)propionic acid (ANP) linker. The modification processes of the Si wafer surface were monitored by Fourier transform infrared spectroscopy by attenuated total reflection (FTIR-ATR) and fast-scanning atomic force microscopy (FS-AFM). The attachment of IgG and its release reaction on the Si surface via the photochemical cleavage of the ANP linker were observed by FS-AFM and an enzyme-linked immunosorbent assay (ELISA). Furthermore, it was possible to selectively collect SP2/O cells that express HEL on their cell membranes (SP2/O-HEL) on the Si wafer device. Photochemical cleavage of the ANP linker facilitated the effective release of living SP2/O cells, whose viability was verified by staining experiments using tripan-blue. Moreover, it was possible to re-culture the recovered cells. This methodology represents an effective strategy for isolating intact target cells in the biological and medicinal sciences and related fields.