Molecular Weight Estimation for Native Proteins Using High-Performance Size-Exclusion Chromatography
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The chromatographic separation of proteins from small molecules on the basis of size was first described by Porath and Flodin, who called the process “gel filtration” (1). Moore applied a similar principle to the separation of polymers on crosslinked polystyrene gels in organic solvents, but named this “gel permeation chromatography” (2). Both terms came to be used by manufacturers of such supports for the separation of proteins, leading to some confusion. The term size-exclusion chromatography is more descriptive of the principle on which separation is based and has largely replaced the older names, although the expression “gel filtration” is still commonly used by biochemists to describe separation of proteins in aqueous mobile phases. Hag el gives a comprehensive review of the subject with emphasis on proteins (3) and there is also much useful information in a technical booklet published by a leading manufacturer (4).
KeywordsBlue Dextran Aqueous Mobile Phasis Dihydrogen Orthophosphate Hydrogen Orthophosphate High Back Pressure
- 2.Moore, J. C. (1964) Gel permeation chromatography. I. A new method for molecular weight distribution of high polymers. J. Polymer Sci. A2, 835–843.Google Scholar
- 3.Hagel, L. (1989) Gel filtration, in Protein Purification: Principles, High Resolution Methods and Applications (Janson, J. C., and Ryden, L., eds.), pp. 63–106. VCH, New York, pp. 63-106.Google Scholar
- 4.Amersham Pharmacia Biotech. 1998. Gel Filtration Principles and Methods, 8th edit., Lund, Sweden.Google Scholar