title: Microaffinity purification of proteins based on photolytic elution: toward an efficient microbead affinity chromatography on a chip.
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 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.
authors.affiliation: School of Chemical Engineering, Seoul National University, Kwanak-Gu, Seoul, Korea.
authors.forename: Woo-Jae
authors.initials: WJ
authors.lastname: Chung
authors.forename: Min-Soo
authors.initials: MS
authors.lastname: Kim
authors.forename: Suhyung
authors.initials: S
authors.lastname: Cho
authors.forename: Sung-Soo
authors.initials: SS
authors.lastname: Park
authors.forename: Jong-Ho
authors.initials: JH
authors.lastname: Kim
authors.forename: Yong-Kweon
authors.initials: YK
authors.lastname: Kim
authors.forename: Byung-Gee
authors.initials: BG
authors.lastname: Kim
authors.forename: Yoon-Sik
authors.initials: YS
authors.lastname: Lee
mesh_terms: D001710:Biotin; D002846:Chromatography, Affinity / Q000379:methods*; D056656:Lab-On-A-Chip Devices; D046208:Microchip Analytical Procedures / Q000379:methods*; D008863:Microspheres; D008904:Miniaturization; D010782:Photolysis; D011506:Proteins / Q000302:isolation & purification*; D012313:RNA; D019809:Streptavidin / Q000302:isolation & purification
chemical_list: D011506:Proteins; D012313:RNA; D001710:Biotin; D019809:Streptavidin