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Capillary electrophoresis is a popular analytical technique that allows the separation of molecules based on their electrophoretic mobility with the use of an applied voltage in a capillary tube containing polymer/gel mediums. The electrophoretic mobility is dependent on the charge of the molecule, the viscosity, and the atom's radius. Capillary electrophoresis can be coupled online with MS to be applied for analysis of proteins and peptides. In the early 1990s, chip-based capillary electrophoresis was designed and developed based on the principle of capillary electrophoresis. Microchip capillary electrophoresis can be integrated with various miniaturized functional elements to realizing mixture, reaction, concentration, and separation of analytes on a chip, greatly decreasing injection volumes, reducing separation lengths, resulting in a reduction of analysis time. As a new and powerful analytical technology, microchip capillary electrophoresis is also applied for high-speed separation of biological compounds such as DNA and proteins.
Fig. 1 Capillary electrophoresis microchip.1
Compared to traditional capillary electrophoresis, chip-based capillary electrophoresis has several advantages, including
Being a silicon compound, glass is an important substrate material in the construction of microchip capillary electrophoresis systems, with the advantages of excellent optical properties, well-developed microfabrication, and the same surface chemistry as the fused silica capillaries used for conventional capillary electrophoresis. However, there are also disadvantages such as complicated processing, high manufacturing cost, and fragility. In view of these disadvantages, polymer substrates for the fabrication of microchips have attracted a significant amount of attention.
Polymer substrates are cheaper and easier to manufacture than glass. The polymer is manufactured through the use of direct-write protocols using electron beams, ultraviolet lasers, or X-rays or through a molding process. Either manufacturing method can provide high-throughput and use lower-cost materials. The main limitation of the polymers used for capillary electrophoresis microchips is that the surface chemistry of these materials is poorly understood and poorly controlled.
Injection, separation, and detection is a standard analysis protocol for microchip capillary electrophoresis.
Microchip capillary electrophoresis technique has been applied for DNA separation for sequencing and fragment analysis. The operation method is very similar to traditional capillary gel electrophoresis.
Microchip capillary electrophoresis can be also applied for protein analysis, involving separation of fluorescently labeled proteins, characterization of proteins, and measuring the purity of protein samples. Especially microchip-based capillary electrophoretic immunoassays allow direct measurement of protein analytes from complex samples such as blood and urine, making them become very useful tools for clinical application.
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Reference
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