Recent Developments in Analytical Science - Electrophoresis

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Education Article

  • Published: Jul 18, 2016
  • Channels: Gas Chromatography / Proteomics & Genomics / Ion Chromatography / Electrophoresis / HPLC / Proteomics / Base Peak / Atomic / MRI Spectroscopy / NMR Knowledge Base / Raman / X-ray Spectrometry / Infrared Spectroscopy

Electrophoresis

Electrophoresis presents different options for separating components before analysis based on the molecular mass of their charged species. One- or two-dimensional gel electrophoresis represents about 60% of the electrophoresis instrumentation market and is a key process in proteomics studies, separating proteins on the basis of their molecular mass for 1D separation or isoelectric point followed by molecular mass for 2D, the latter option providing better separation but taking longer.

In 2D differential gel electrophoresis, the proteins on different gels are compared using fluorescent dyes, so that the proteins from a diseased subject can be compared to those in a healthy subject. Despite its popularity, 2DE has a poor reproducibility and a narrow dynamic range, although the latter can be improved by prior removal of the most abundant proteins like albumin, or by prefractionation and protein enrichment.47 Better protein separation can also be achieved by adding ionic liquids to the buffer solutions.

Although it is a slow process, gel electrophoresis can be accelerated by miniaturisation, one proposed system using disposable cassettes containing the gel with conducting polymers instead of metal electrodes. The developers declared that the system “has the potential to pave the way for the development of low-cost, disposable, miniaturized GE systems for accelerated analysis in areas such as proteomics and medical diagnosis.48

Another major type of electrophoresis is capillary electrophoresis, which has also experienced the advantages of miniaturisation with the development of portable and microchip systems over the last decade and that are seen as a key route to point-of-care diagnostics and monitoring. It is also a key technique for DNA analysis and has been employed heavily in the clinical setting, drug metabolism studies and environmental analysis.

2D Capillary electrophoresis procedures have been developed to provide better resolution and peak capacity. They involve several modes, such as capillary isoelectric focussing/capillary electrophoresis and capillary sieving electrophoresis/micellar electrokinetic capillary chromatography with many different types of interface in conventional and microchip systems. 3D separations are also possible.49

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