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

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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.