Fully Automated Two-Dimensional Electrophoresis System for High-Throughput Protein Analysis

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.