Selective capture and collection of live target cells using a photoreactive silicon wafer device modified with antibodies via a photocleavable linker.
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.
Center for Technologies against Cancer, Tokyo University of Science, 2641 Yamazaki, Noda, Japan.
Shinya
S
Ariyasu
Kengo
K
Hanaya
Eita
E
Watanabe
Toshihiro
T
Suzuki
Kazutaka
K
Horie
Masanori
M
Hayase
Ryo
R
Abe
Shin
S
Aoki
D000818:Animals; D000906:Antibodies / Q000737:chemistry* / Q000276:immunology; D000937:Antigen-Antibody Reactions; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002470:Cell Survival; D002645:Chickens; D004912:Erythrocytes / Q000166:cytology* / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation; D009113:Muramidase / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D009596:Nitrophenols / Q000138:chemical synthesis / Q000737:chemistry*; D055668:Photochemical Processes; D011422:Propionates / Q000138:chemical synthesis / Q000737:chemistry*; D012825:Silicon / Q000737:chemistry*
C107988:3-amino-3-(2-nitrophenyl)propionic acid; D000906:Antibodies; D007074:Immunoglobulin G; D009596:Nitrophenols; D011422:Propionates; C421864:hen egg lysozyme; D009113:Muramidase; D012825:Silicon