We describe a miniaturized instrument capable of performing 2-DE. Our miniaturized device is able to perform IEF and polyacrylamide slab gel electrophoresis (PASGE) in the same unit. It consists of a compartment for a first-dimensional IEF gel, which is connected to a second-dimensional PASGE gel. The focused samples are automatically transferred from the IEF gel to the PASGE gel by electromigration. Our preliminary experiments show that the device is able to focus and separate a mixture of proteins in approximately 1 h, excluding the time required for the staining procedure. On average, the gel-to-gel retardation factor (Rf) variation was 6.2% (+/-0.9%) and pI variation was 2.5% (+/-0.6%). Separated protein spots were excised from stained gels, digested with trypsin, and further identified by MS, thus enabling direct proteomic analysis of the separated proteins.
The present article describes a miniaturized polyacrylamide slab gel electrophoresis-chip (PASGE-Chip) that can rapidly separate a set of predefined samples as well as cell lysate samples for clinical diagnosis. The chip consists of a polymethyl methacrylate (PMMA) upper unit (25 x 30 x 10 mm, width x length x depth) with integrated buffer chambers, running electrodes and loading wells and a bottom unit comprising a silicon dioxide-coated silicon plate with embossed gel chamber (11 x 15 x 0.37 mm). This miniaturized device was designed to be fast, easy to use and cheap to produce. The polyacrylamide slab gel electrophoresis can be performed in less than 10 min with low voltage. The gel-to-gel repeatability is around 3.8%. The limit of detection is approx. 10 ng as determined by Coomassie staining of selected standard proteins, and corresponds to a 10-fold increase in sensitivity as compared with a common size PAGE analysis device (e.g. 10 x 7 cm). The device was successfully applied to peptide mass fingerprint analysis, protein sequencing and ultra-sensitive immunodetection, and the performance was compared to a commonly used regular PAGE device.
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