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

利用光可裂解接头捕获和收集循环肿瘤细胞方法的开发

• 批准号: 22390005
• 负责人: AOKI Shin
• 金额: $ 11.98万
• 依托单位: Tokyo University of Science
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22390005/
• 关键词: 血中循環腫瘍細胞; 光分解反応; がん診断; シリコン基板; 抗体

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.

循环肿瘤细胞（CTC）已被定义为在外周血中发现的实体瘤来源的癌细胞。一般认为，这些CTC从患者的原发性肿瘤中分离，进入血液，在体内四处游走，然后附着到远处组织。因此，一般认为CTC的定量检测和分析对于癌症患者的个人诊断是有用的。在这项工作中，开发了一种用于捕获和回收活靶细胞的装置。平台是通过可光裂解的3-氨基-3-（2-硝基苯基）丙酸（ANP）接头用抗HEL抗体（抗HEL-IgG，HEL=鸡蛋溶菌酶）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）对硅片表面改性过程进行了监测。通过FS-AFM和酶联免疫吸附测定（ELISA）观察了IgG在Si表面的附着及其通过ANP连接子的光化学切割而释放的反应。此外，可以在Si晶片装置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连接子的光化学裂解促进了活SP2/O细胞的有效释放，其活力通过使用三泛蓝的染色实验来验证。此外，可以再培养回收的细胞。该方法代表了在生物和医学科学及相关领域中分离完整靶细胞的有效策略。

项目成果

pH変化に応答するシクロメタレート型イリジウム(III)錯体の設計と合成

响应pH变化的环金属酸盐型铱(III)配合物的设计与合成

• 发表时间: 2011
• 作者: 諸見里真輔;久松洋介;大和田紘喜;松生泰樹;青木伸
• 通讯作者: 青木伸

Design and Synthesis of a Photocleavable Linker for the Isolation Device of Circulating Tumor Cells

用于循环肿瘤细胞分离装置的光裂解接头的设计与合成

• 发表时间: 2011
• 作者: Shinya Ariyasu;Kengo Hanaya;Megumi Tsunoda;Misato Hoshi;Masanori Kitamura;Masanori Hayase;Ryo Abe;Shin Aoki
• 通讯作者: Shin Aoki

One-pot Chemoenzymatic Synthesis of Optically Active Compounds Using Chiral Zinc(II) Complexes and Enzymes in Aqueous Solvent

在水溶剂中使用手性锌 (II) 配合物和酶一锅化学酶法合成光学活性化合物

• 发表时间: 2011
• 作者: Shin Aoki;Shotaro Sonoike;Toshinari Itakura;Susumu Itoh;Masanori Kitamura
• 通讯作者: Masanori Kitamura

血中腫瘍細胞捕捉のための寸法によるファーストスクリーニング

捕获血液中肿瘤细胞的三维首次筛选

• 发表时间: 2011
• 作者: 陶山敬樹;有安真也;花屋賢悟;青木伸;鈴木利宙;安部良;早瀬仁則
• 通讯作者: 早瀬仁則

微小流路を用いたシリコン基板表面上の細胞付着力測定-光解離性リンカーによる付着力の低下-

使用微通道测量硅基底表面上的细胞粘附力 - 由于光可裂解连接体而降低粘附力 -

• 发表时间: 2012
• 作者: 陶山敬樹;有安真也;花屋賢悟;青木伸;鈴木利宙;安部良;早瀬仁則
• 通讯作者: 早瀬仁則