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Microaffinity purification of proteins based on photolytic elution: Toward an efficient microbead affinity chromatography on a chip

Title
Microaffinity purification of proteins based on photolytic elution: Toward an efficient microbead affinity chromatography on a chip
Author
김종호
Keywords
microaffinity purification; microbead affinity chromatography; miniaturization; photo-cleavable ligand; photolytic elution
Issue Date
2005-02
Publisher
WILEY-V C H VERLAG GMBH
Citation
ELECTROPHORESIS, v. 26, No. 3, Page. 694-702
Abstract
A bead affinity chromatography system, which was based on the photolytic elution method, was integrated into a glass-silicon microchip to purify specific target proteins. CutiCore(R) beads, which were coupled with a photo-cleavable ligand, such as biotin and an RNA aptamer, were introduced into a filter chamber in the microchip. The protein mixture containing target protein labeled with fluorescein isothiocyanate (FITC) was then passed through the packed affinity beads in the microchamber by pressure-driven flow. During the process, the adsorbed protein on the bead was monitored by fluorescence. The concentrated target protein on the affinity bead was released by simple irradiation with UV light at a wavelength of 360 nm, and subsequently eluted with the phosphate buffer flow. The eluted target protein was quantitatively detected via the fluorescence intensity measurements at the downstream of the capillary connected to the outlet of the microchip. The microaffinity purification allowed for a successful method for the identification of specific target proteins from a protein mixture. In addition, the feasibility of this system for use as a diagnosis chip was demonstrated.
URI
https://onlinelibrary.wiley.com/doi/abs/10.1002/elps.200410005https://repository.hanyang.ac.kr/handle/20.500.11754/110221
ISSN
0173-0835; 1522-2683
DOI
10.1002/elps.200410005
Appears in Collections:
COLLEGE OF ENGINEERING SCIENCES[E](공학대학) > MATERIALS SCIENCE AND CHEMICAL ENGINEERING(재료화학공학과) > Articles
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