Capillary zone electrophoresis of proteins in semidilute polymer solutions: Inter- and intra-polymer predictability of size-dependent retardation

Št́astná, Miroslava; Radko, Sergey P.; Chrambach, Andreas

doi:10.1002/(sici)1522-2683(19991001)20:14<2884::aid-elps2884>3.0.co;2-5

Cited by 6 publications

(7 citation statements)

References 22 publications

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“…It has been reported that the limiting value of the protein radius is 2 nm in applying theories of protein retardation in some polymer solutions. However, it has also been emphasized that protein retardation seems to be polymer-specific rather than universal 3 Ferguson plot analysis of the mobility of three charged isoforms of carbonic anhydrase (CA1−CA3) and a sample of HSA.…”

Section: Resultsmentioning

confidence: 99%

“…Effect of Mg 2+ on Protein Retardation in PVP Solutions. The effect of cross-linking agents cannot be explained using a simplified equation relating protein retardation in semidilute polymer solutions during capillary electrophoresis . A simplified equation suggests that protein retardation in a polymer solution is related only to the polymer concentration c : where μ/μ 0 is the retardation, expressed as the ratio between mobility in the polymer solution and in the free solution.…”

Section: Resultsmentioning

confidence: 99%

“…Despite the advantages of low-viscosity polymers for DNA separation, little is known about retardation of proteins in these types of polymers. For example, long DNA molecules can change their conformation and reptate to find a path through a polymer network during electrophoresis, whereas a nondenatured protein molecule normally retains its structure and therefore moves as a particle having a fairly constant radius . In a solution of linear polymers, when a particle is larger than the pores within the polymer network, the network will adjust to enlarge the local mesh size, with entangled chains sliding from one another, and this movement is detrimental to the resolution of the separation .…”

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confidence: 99%

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Dynamic Cross-Linking Effect of Mg²⁺ To Enhance Sieving Properties of Low-Viscosity Poly(vinylpyrrolidone) Solutions for Microchip Electrophoresis of Proteins

et al. 2007

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We report a dynamic cross-linking effect of Mg2+ that enhances the sieving properties of low-viscosity poly(vinylpyrrolidone) (PVP) solutions. A low-viscosity PVP solution was applied to nondenaturing microchip electrophoresis of protein samples using microchips made of poly(methyl methacrylate). The separation resolution of nondenatured protein markers in 1.8% PVP solution was improved by adding 1-20 mM MgCl2. We studied the effect of the ratio of cross-linking agent on mobility of protein samples and showed that protein retardation (ln micro/micro0) is correlated with the ratio of cross-linking agent to PVP ([cMg2+/cPVP]) as ln micro/micro0=A'[cMg2+/cPVP]b'. A' was related to the protein radius (R), and b' was found to be 0.72 for proteins with R=2.4 nm and 0.82 for proteins with R=1.85 nm. A structural study of PVP in semidilute solutions using dynamic light scattering showed that incremental increases of Mg2+ ion concentration from 5 to 20 mM in 1.8% PVP solution increased the hydrodynamic radius of PVP polymers by 20%.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

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Dynamic Cross-Linking Effect of Mg²⁺ To Enhance Sieving Properties of Low-Viscosity Poly(vinylpyrrolidone) Solutions for Microchip Electrophoresis of Proteins

et al. 2007

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“…The success of DNA separation using PEO containing GNPs opens the avenue of using nanomaterials for the separation of biomolecules. We point out that the importance of nanomaterials for bioanalysis can be further realized once biofunctional nanomaterials that are highly specific to proteins and DNA of interest are prepared [214][215][216]. …”

Section: Discussionmentioning

confidence: 99%

Analysis of Nucleic Acids and Proteins in Capillary Electrophoresis and Microchip Capillary Electrophoresis Using Polymers as Additives of the Background Electrolytes

Hsieh¹,

Chiu²,

Tseng³

et al. 2006

CAC

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This review article deals with recent development in the analyses of proteins and DNA by capillary electrophoresis (CE) and microchip CE (MCE) using polymer solutions. Polymer solutions are commonly prepared from linear polymers such as linear polyacrylamide, poly(ethylene oxide), hydroxyethyl cellulose, and poly(vinylpyrrolidone), which possess the advantages of low fluorescence background, self-coating capability, and high sieving ability. We discuss the key parameters that affect the separation efficiency, including electroosmotic flow, chemical and physical properties of polymers, background electrolytes, and additives. The advantages and disadvantages of CE and MCE in conjunction with laser-induced fluorescence using polymer solutions for the analyses of DNA and proteins in biological fluids are highlighted. Several recently developed CE approaches such as gradient techniques and sample stacking for efficient and sensitive analyses of DNA and proteins are emphasized. We also address the trend of using nanomaterials for bioanalysis in CE and MCE.

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“…Dextran is a branched polysaccharide and thus even dextran‐based oligosaccharides can provide effective size‐dependent separations 28, 29. From synthetic polymers, poly(ethylene oxide) (PEO) 24, 30–33 and poly(vinyl alcohol) 25, 30, 34 were mostly studied. Some less common gel matrices have been reviewed by Mikšík et al .…”

Section: Introductionmentioning

confidence: 99%

Pluronic F‐127 as the buffer additive in capillary entangled polymer electrophoresis: Some fundamental aspects

Křížek

Coufal

Tesařová

et al. 2010

J of Separation Science

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Polymeric macromolecules of well-designed structures and specific properties open promising directions in the capillary entangled polymer electrophoresis. Pluronic F-127, as a thermoassociating polymer, possesses some unique properties that can be utilized in capillary entangled polymer electrophoresis of amino acids, peptides and proteins. In this study, we studied properties of Pluronic F-127 polymer as an additive to BGE for the separation of peptides and proteins. The influence of the thermoassociation on separation selectivity was studied. The addition of Pluronic caused severe instabilities of the electrical current and the signal of the UV detector. This study reveals remarkable positive effect of a low pressure applied to the inlet buffer vial during the analysis, which apparently stabilizes the electrical current and the detector signal. The effect of hydrodynamic flow induced by the pressure applied on the separation efficiency was studied and the significance of this effect was discussed. Pluronic F-127, as a representative of synthetic macromolecules, was compared with dextran, as a representative of natural polymers, in terms of separation power, selectivity and repeatability of migration times.

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Capillary zone electrophoresis of proteins in semidilute polymer solutions: Inter- and intra-polymer predictability of size-dependent retardation

Cited by 6 publications

References 22 publications

Dynamic Cross-Linking Effect of Mg²⁺ To Enhance Sieving Properties of Low-Viscosity Poly(vinylpyrrolidone) Solutions for Microchip Electrophoresis of Proteins

Dynamic Cross-Linking Effect of Mg²⁺ To Enhance Sieving Properties of Low-Viscosity Poly(vinylpyrrolidone) Solutions for Microchip Electrophoresis of Proteins

Analysis of Nucleic Acids and Proteins in Capillary Electrophoresis and Microchip Capillary Electrophoresis Using Polymers as Additives of the Background Electrolytes

Pluronic F‐127 as the buffer additive in capillary entangled polymer electrophoresis: Some fundamental aspects

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Capillary zone electrophoresis of proteins in semidilute polymer solutions: Inter- and intra-polymer predictability of size-dependent retardation

Cited by 6 publications

References 22 publications

Dynamic Cross-Linking Effect of Mg2+ To Enhance Sieving Properties of Low-Viscosity Poly(vinylpyrrolidone) Solutions for Microchip Electrophoresis of Proteins

Dynamic Cross-Linking Effect of Mg2+ To Enhance Sieving Properties of Low-Viscosity Poly(vinylpyrrolidone) Solutions for Microchip Electrophoresis of Proteins

Analysis of Nucleic Acids and Proteins in Capillary Electrophoresis and Microchip Capillary Electrophoresis Using Polymers as Additives of the Background Electrolytes

Pluronic F‐127 as the buffer additive in capillary entangled polymer electrophoresis: Some fundamental aspects

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Product

Resources

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Dynamic Cross-Linking Effect of Mg²⁺ To Enhance Sieving Properties of Low-Viscosity Poly(vinylpyrrolidone) Solutions for Microchip Electrophoresis of Proteins

Dynamic Cross-Linking Effect of Mg²⁺ To Enhance Sieving Properties of Low-Viscosity Poly(vinylpyrrolidone) Solutions for Microchip Electrophoresis of Proteins