Influence of ignored and well-known zone distortions on the separation performance of proteins in capillary free zone electrophoresis with special reference to analysis in polyacrylamide-coated fused silica capillaries in various buffers

Mohabbati, Sheila; Westerlund, Douglas

doi:10.1016/j.chroma.2004.08.073

Cited by 9 publications

(36 citation statements)

References 67 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The decrease in sensitivity can be explained by a possible loss of peptides during the electrokinetic injection given that the stacking process should occur at the extremity of the capillary. Indeed, it has recently been shown that protein losses due to sedimentation of the enriched protein zone might occur during electrokinetic injection under zonesharpening conditions [28].…”

Section: Eksmentioning

confidence: 99%

Electrokinetic supercharging for highly efficient peptide preconcentration in capillary zone electrophoresis

2008

View full text Add to dashboard Cite

Electrokinetic supercharging has been integrated in CZE for the development of a highly sensitive methodology for protein tryptic digest analysis. A careful choice of the experimental conditions led to sensitivity enhancement factors between 1000 and 10 000 whilst maintaining a satisfactory resolution. Peptides in the low nanomolar concentration range have been detected despite the use of the poorly sensitive UV absorbance detection mode. The buffer system used in this study is fully suitable for coupling CE to MS.

show abstract

Section: Eksmentioning

confidence: 99%

Electrokinetic supercharging for highly efficient peptide preconcentration in capillary zone electrophoresis

2008

View full text Add to dashboard Cite

show abstract

“…Also, dynamic capillary coating is simple, but may complicate the detection, for example, when MS is used. The preparation of permanently coated capillaries is somewhat cumbersome and prone to variability, but for a given capillary usually results in stable performance for quite a long time [5][6][7][8]. However, adsorption problems may be encountered even with coated capillaries [6,7].…”

mentioning

confidence: 99%

Utilizing the pH hysteresis effect for versatile and simple electrophoretic analysis of proteins in bare fused-silica capillaries

Bohlin¹,

Blomberg²,

Heegaard³

2005

Electrophoresis

View full text Add to dashboard Cite

The analysis of peptides and proteins by CE is often desirable due to low sample consumption and possibilities for nondenaturing yet highly effective separations. However, adsorption to the inner surfaces of fused-silica capillaries often is detrimental to such analyses. This phenomenon is especially pronounced in the analysis of basic proteins and proteins containing exposed positively charged patches. To avoid wall interactions numerous buffer additives and static and dynamic wall coating principles have been devised. We previously showed (J. Chromatogr. A 2004, 1059, 215-222) that CE of the basic protein beta2-glycoprotein was rendered possible by an acidic pretreatment step, and we attributed this observation to the so-called pH hysteresis effect that influences the time for pH equilibration of the capillary wall and thus the effective wall charge and the electroosmotic mobility. We here investigate the effects of different pretreatment techniques on EOF values and on the rate of the deprotonation of silanol groups when performing the electrophoresis at neutral pH. We show the utility of this simple approach for the CE analysis of a number of basic proteins in plain silica capillaries at physiological pH.

show abstract

“…Linear polyacrylamide has been used to eliminate EOF in both fused-silica capillary [74][75][76] and channels in PDMS microchips [77]. Stability of the polyacrylamide coating was enhanced by cross-linking [78].…”

Section: Static Wall Coatingmentioning

confidence: 99%

“…However, the composition of BGE affects separation efficiency, too. Several BGEs for CZE of proteins have been compared and 120 mM ammonium acetate and 150 mM ammonium hydroxyacetate, pH 4 were selected as the most promising buffers for CZE of proteins [74,75].…”

Section: Improving Separationmentioning

confidence: 99%

Capillary electrophoresis of proteins 2003–2005

Dolnı́k¹

2006

Electrophoresis

139

117

View full text Add to dashboard Cite

This review article with 304 references describes recent developments in CE of proteins, and covers the two years since the previous review (Hutterer, K., Dolník, V., Electrophoresis 2003, 24, 3998-4012) through Spring 2005. It covers topics related to CE of proteins, including modeling of the electrophoretic migration of proteins, sample pretreatment, wall coatings, improving separation, various forms of detection, special electrophoretic techniques such as affinity CE, CIEF, and applications of CE to the analysis of proteins in real-world samples including human body fluids, food and agricultural samples, protein pharmaceuticals, and recombinant protein preparations.

show abstract

Influence of ignored and well-known zone distortions on the separation performance of proteins in capillary free zone electrophoresis with special reference to analysis in polyacrylamide-coated fused silica capillaries in various buffers

Cited by 9 publications

References 67 publications

Electrokinetic supercharging for highly efficient peptide preconcentration in capillary zone electrophoresis

Electrokinetic supercharging for highly efficient peptide preconcentration in capillary zone electrophoresis

Utilizing the pH hysteresis effect for versatile and simple electrophoretic analysis of proteins in bare fused-silica capillaries

Capillary electrophoresis of proteins 2003–2005

Contact Info

Product

Resources

About