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Capillary electrophoresis separates molecules based on size and electrical charge. An electric field is applied, causing molecules to move at different speeds. UV light or fluorescent detection is used to locate molecules. It has applications in genetics, food science, and medical research.
In biochemistry, capillary electrophoresis is a technique used to separate molecules based on their size and electrical charge. It is also often called capillary zone electrophoresis or capillary gel electrophoresis. Capillary electrophoresis analysis is based on the principle that molecules possess different electric charges and different weights. Therefore, when molecules in a substrate are exposed to an electric field, different molecules will move in different directions and at different speeds within the substrate.
When an electric field is applied to a substrate by means of an electrode at either end of the field, positively charged molecules will move towards the negative electrode and negatively charged molecules will move towards the positive electrode. The relative speed at which the molecule will move through the substrate is determined by a characteristic known as the hydrodynamic size of the molecule. The hydrodynamic size of a molecule depends on two factors, its mass and the strength of its positive or negative charge. A large molecule with a strong positive charge will tend to move relatively rapidly towards the negative electrode in an electric field. A small molecule with a weak negative charge will tend to move much more slowly towards the positive electrode in an electric field.
Once a charge has been applied to a sample for a specified amount of time and the molecules to be analyzed have been separated, it is necessary to show the scientist where the different molecules are located in the substrate. In many modern capillary electrophoresis systems this is achieved with ultraviolet (UV) light. UV illumination can show the user the physical path that has been taken by a molecule, or set of molecules, through the substrate. In other systems, the molecules to be analyzed are first treated with chemicals to give them a fluorescent property. Fluorescent detection methods can be very sensitive, but they are not suitable for every type of molecule.
The applications of capillary electrophoresis in science are many and varied. Capillary electrophoresis sequencing is a process, especially used by geneticists, in which capillary electrophoresis is used to analyze molecules of the genetic material Deoxyribo Nucleic Acid (DNA). Food scientists can use capillary electrophoresis protein analysis to determine different characteristics of the protein content of foods. In medical research, this process has many applications, such as analyzing antibodies and studying how they bind to other molecules.
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