title: Fully automated two-dimensional electrophoresis system for high-throughput protein analysis.
abstract: We developed a fully automated electrophoresis system for rapid and highly reproducible protein analysis. All the two-dimensional (2D) electrophoresis procedures including isoelectric focusing (IEF), on-part protein staining, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and in situ protein detection were automatically completed. The system comprised Peltiert devices, high-voltage generating devices, electrodes, and three disposable polymethylmethacrylate (PMMA) parts for IEF, reaction chambers, and SDS-PAGE. Because of miniaturization of the IEF part, rapid IEF was achieved in 30 min. A gel with a tapered edge gel on the SDS-PAGE part realized a connection between the parts without use of a gluing material. A biaxial conveyer was employed for the part relocation, sample introduction, and washing processes to realize a low-maintenance and cost-effective automation system. Performances of the system and a commercial minigel system were compared in terms of detected number, resolution, and reproducibility of the protein spots. The system achieved high-resolution comparable to the minigel system despite shorter focusing time and smaller part dimensions. The resulting reproducibility was better or comparable to the performance of the minigel system. Complete 2D separation was achieved within 1.5 h. The system is practical, portable, and has automation capabilities.
authors.affiliation: Research Center of Advanced Bionics, National Institute of Advanced Industrial Science and Technology, c/o Katayanagi Institute, Tokyo University of Technology, 1404-1, Katakura, Hachioji, Tokyo 192-0982, Japan.
authors.forename: Atsunori
authors.initials: A
authors.lastname: Hiratsuka
authors.forename: Hideki
authors.initials: H
authors.lastname: Kinoshita
authors.forename: Yuji
authors.initials: Y
authors.lastname: Maruo
authors.forename: Katsuyoshi
authors.initials: K
authors.lastname: Takahashi
authors.forename: Satonari
authors.initials: S
authors.lastname: Akutsu
authors.forename: Chie
authors.initials: C
authors.lastname: Hayashida
authors.forename: Koji
authors.initials: K
authors.lastname: Sakairi
authors.forename: Keisuke
authors.initials: K
authors.lastname: Usui
authors.forename: Kisho
authors.initials: K
authors.lastname: Shiseki
authors.forename: Hajime
authors.initials: H
authors.lastname: Inamochi
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Nakada
authors.forename: Kouhei
authors.initials: K
authors.lastname: Yodoya
authors.forename: Ichiji
authors.initials: I
authors.lastname: Namatame
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Unuma
authors.forename: Makoto
authors.initials: M
authors.lastname: Nakamura
authors.forename: Kosuke
authors.initials: K
authors.lastname: Ueyama
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Ishii
authors.forename: Kazuyoshi
authors.initials: K
authors.lastname: Yano
authors.forename: Kenji
authors.initials: K
authors.lastname: Yokoyama
mesh_terms: D001331:Automation; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D007525:Isoelectric Focusing / Q000379:methods; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D015203:Reproducibility of Results; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D013194:Staining and Labeling
chemical_list: D011506:Proteins; D012967:Sodium Dodecyl Sulfate