Estimation of Hydrodynamic Volume of Proteins Using High-Performance Size-Exclusion Chromatography and Intrinsic Viscosity Measurement: An Attempt At Universal Calibration

Chikazumi, Nobutoshi; Ohta, Takao

doi:10.1080/01483919108049261

Cited by 11 publications

(2 citation statements)

References 45 publications

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“…By contrast, the elution of the large SDS-protein complexes follows a different curve, being characterized by a relatively large R^ for the same elution volume. The data on GuHCl denatured and native proteins are in agreement with data reported by others (8,15), and by Nave et al (18) for TSK 6000 PW, but not for Superose 6 columns. Horiike et al (8) also showed that the agreement is less satisfactory if a frictional coefficient based Stokes radius is used.…”

Section: The Use Of Gel Chromatography For Characterization Of Randomsupporting

confidence: 93%

“…conformation (13), we found no systematic difference between R § and in a plot relating these para meters and molecular masses (14). Values of R^ for proteins can be found in Refs 8,9,[15][16][17] for an updated summary see Nave et al (18). It has been pointed out that the elution position of proteins is better correlated with the viscosity-based Stokes radius than with R s , as calculated from the frictional coefficient.…”

Section: The Use Of Gel Chromatography For Characterization Of Randommentioning

confidence: 99%

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Gel Chromatography as an Analytical Tool for Characterization of Size and Molecular Mass of Proteins

Maire

Ghazi

Møller

1996

ACS Symposium Series

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Although experience with watersoluble, globular proteins supports Stokes radius as an appropriate size parameter, governing elution position, for both classical and HPLC gel columns, extrapolation of this concept to other kinds of conformation and substances requires caution. Current evidence suggests the viscosity based Stokes radius as the best suited size parameter for many flexible macromolecules (guanidinium HCl denatured proteins, dextrans), whereas many detergent micelles and detergent solubilized membrane proteins elute somewhat earlier, and elongated (e.g. myosin) or SDS denatured proteins elute later, than indicated by their hydrodynamic behaviour. The review includes a discussion of the use of globular proteins of known size to characterize gel pore size distribution.The purpose of this review is to briefly consider the scope of gel chromatography, or size exclusion chromatography (SEC), in the determination of sizes and molecular masses of proteins. This kind of separation is performed either as classical type chromatography, using gels such as Sepharose (agarose), Sephacryl (allyl-dextran crosslinked with Ν,Ν-methylene bisacrylamide) and Superose, or by HPLC with the aid of matrices capable of withstanding high pressures such as silica based TSK SW-or polyacrylamide based TSK PW columns. We first briefly review the theoretical foundation for use of the technique in the estimation of molecular mass and size of watersoluble, globular proteins. We then proceed to consider the use of the technique to analyze other kinds of proteins (elongated or fibrous type proteins, detergent-solubilized membrane proteins) and the effect of a disordered conformation (SDS-or GuHCl denaturation). Finally, we consider the use of globular proteins in the characterization of the pore characteristics of the gels.

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Section: The Use Of Gel Chromatography For Characterization Of Randomsupporting

confidence: 93%

Section: The Use Of Gel Chromatography For Characterization Of Randommentioning

confidence: 99%

Gel Chromatography as an Analytical Tool for Characterization of Size and Molecular Mass of Proteins

Maire

Ghazi

Møller

1996

ACS Symposium Series

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Universal calibration of size-exclusion chromatography for proteins in guanidinium hydrochloride including the high-molecular-mass proteins titin and nebulin

Nave

Weber

Potschka

1993

Journal of Chromatography A

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Factorial experimental design and principal component analysis of the interaction of animal glues with polymeric and silica-based stationary phases in size exclusion chromatography

Weatherell

Lai

1994

Journal of Chromatography A

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Estimation of Hydrodynamic Volume of Proteins Using High-Performance Size-Exclusion Chromatography and Intrinsic Viscosity Measurement: An Attempt At Universal Calibration

Cited by 11 publications

References 45 publications

Gel Chromatography as an Analytical Tool for Characterization of Size and Molecular Mass of Proteins

Gel Chromatography as an Analytical Tool for Characterization of Size and Molecular Mass of Proteins

Universal calibration of size-exclusion chromatography for proteins in guanidinium hydrochloride including the high-molecular-mass proteins titin and nebulin

Factorial experimental design and principal component analysis of the interaction of animal glues with polymeric and silica-based stationary phases in size exclusion chromatography

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