Abstract
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).
Keywords
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Porath, J. and Flodin, P. (1959) Gel filtration: a method for desalting and group separation. Nature 183, 1657–1659.
Moore, J. C. (1964) Gel permeation chromatography. I. A new method for molecular weight distribution of high polymers. J. Polymer Sci. A2, 835–843.
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.
Amersham Pharmacia Biotech. 1998. Gel Filtration Principles and Methods, 8th edit., Lund, Sweden.
Potschka, M. (1987) Universal calibration of gel permeation chromatography and determination of molecular shape in solution. Analyt. Biochem. 162, 47–64.
Dubin, P. L. and Principi, J. M. (1989) Failure of universal calibration for size exclusion chromatography of rodlike macromolecules versus random coils and globular proteins. Macromolecules 22, 1891–1896.
Ui, N. (1979) Rapid estimation of molecular weights of protein polypeptide chains using high-pressure liquid chromatography in 6 M guanidine hydrochloride. Analyt. Biochem. 97, 65–71.
Kato, Y., Komiya, K., Sasaki, H., and Hashimoto, T. (1980) High-speed gel filtration of proteins in 6 M guanidine hydrochloride on TSK-GEL SW columns. J. Chromatogr. 193, 458–463.
Josic, D., Baumann, H., and Reutter, W. (1984) Size-exclusion high-performance liquid chromatography and sodium dodecyl sulphate-polyacrylamide gel electrophoresis of proteins: a comparison, Analyt. Biochem. 142, 473–479.
Anspach, B., Gierlich, H. U., and Unger, K. K. (1988) Comparative study of Zorbax Bio Series GF 250 and GF 450 and TSK-GEL 3000 SW and SWXL columns in size-exclusion chromatography of proteins. J. Chromatogr. 443, 45–54.
Irvine, G. B. (1987) High-performance size-exclusion chromatography of polypeptides on a TSK G2000SW column in acidic mobile phases. J. Chromatogr. 404, 215–222.
Regnier, F. E. (1983) High performance liquid chromatography of proteins. Meth. Enzymol. 91, 137–192.
Ackers, G. K. (1970) Analytical gel chromatography of proteins. Adv. Protein Chem. 24, 343–446.
Brooks, D. E., Haynes, C. A., Hritcu, D., Steels, B. M., and Muller, W. (2000) Size exclusion chromatography does not require pores. Proc. Natl. Acad. Sci. USA 97, 7064–7067.
Kato, Y., Komiya, K., Sasaki, H., and Hashimoto, T. (1980) Separation range and separation efficiency in high-speed gel filtration on TSK-GEL SW columns. J. Chromatogr. 190, 297–303.
Himmel, M. E. and Squire, P. G. (1981) High pressure gel permeation chromatography of native proteins on TSK-SW columns. Int. J. Peptide Protein Res. 17, 365–373.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2002 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Brent Irvine, G. (2002). Molecular Weight Estimation for Native Proteins Using High-Performance Size-Exclusion Chromatography. In: Walker, J.M. (eds) The Protein Protocols Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1385/1-59259-169-8:573
Download citation
DOI: https://doi.org/10.1385/1-59259-169-8:573
Publisher Name: Humana Press
Print ISBN: 978-0-89603-940-7
Online ISBN: 978-1-59259-169-5
eBook Packages: Springer Book Archive