Selective Capture and Collection of Live Target Cells Using a Photoreactive Silicon Wafer Device Modified with Antibodies via a Photocleavable Linker
Selective Capture and Collection of Live Target Cells Using a Photoreactive Silicon Wafer Device Modified with Antibodies via a Photocleavable Linker
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DOI：
10.1021/la302393p
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发表时间：
2012-09-11
期刊：
影响因子：
3.9
通讯作者：
Aoki, Shin
中科院分区：
文献类型：
--
作者：
Ariyasu, Shinya;Hanaya, Kengo;Aoki, Shin
描述了一种用于捕获和再收集活靶细胞的装置。该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。利用傅里叶变换红外光谱衰减全反射（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直接观察到IgG在Si表面的附着及其通过ANP连接体的光化学切割的释放反应。酶联免疫吸附试验（ELISA）的结果表明，从Si表面的抗HEL-IgG与第二抗体-碱性磷酸酶杂合物（第二IgG-AP）的复合物的光释放发生最小的损害。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）。光化学裂解的ANP接头促进有效释放的活SP2/O细胞，其活力通过使用三泛蓝染色实验验证。此外，可以再培养回收的细胞。这种方法代表了生物和医学科学及相关领域分离完整靶细胞的有效策略。
A device for the capture and recollection of live target cells is described. 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 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 directly by FS-AFM. The results of an enzyme-linked immunosorbent assay (ELISA) indicated that the photorelease of the complex of anti-HEL-IgG with the secondary antibody-alkaline phosphatase hybrid (secondary IgG-AP) from the Si surface occurs with minimum damage. Furthermore, it was possible to collect SP2/O cells selectively 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 reculture the recovered cells. This methodology represents an effective strategy for isolating intact target cells in the biological and medicinal sciences and related fields.