An Improvement of the On-line Electrophoretic Concentration Method for Capillary Electrophoresis of Proteins and Experimental Factors Affecting the Concentration Effect

Wu, Xing-Zheng; Kasashima, Takanori

doi:10.2116/analsci.16.329

Cited by 19 publications

(13 citation statements)

References 18 publications

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“…The CE instrument was similar to that used in our previous works [31,32]. A high-voltage power supply (Matsusada Precision, Osaka, Japan) provided injection and separation voltages.…”

Section: Capillary Electrophoresis Instrumentationmentioning

confidence: 99%

“…Etching of the capillary wall with HF has been reported [32,33]. In this work, we improved the HF etching method.…”

Section: Etching Of the Capillary Wallmentioning

confidence: 99%

“…Other electrophoretic concentration methods, such as sweeping [28], pH junction [29], and the recently reported nonuniform capillary modulation [30], are also based on the velocity difference when ions move across the boundary of a sample plug and a running buffer. A semipermeable microdialysis hollow fiber directly connected to the inlet end of a capillary can also be used for concentration of protein samples since large proteins cannot pass through the hollow fiber [31,32]. Recently, this concentration principle was further developed by etching a short section of capillary into a porous joint which allows electrical conductivity but blocks the analyte ions [33].…”

Section: Introductionmentioning

confidence: 99%

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Isoelectric focusing sample injection for capillary electrophoresis of proteins

Wu¹,

Zhang²,

Onoda³

2005

Electrophoresis

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Carrier ampholyte-free isoelectric focusing (IEF) sample injection (concentration) for capillary electrophoresis (CE) is realized in a single capillary. A short section of porous capillary wall was made near the injection end of a capillary by HF etching. In the etching process, an electric voltage was applied across the etching capillary wall and electric current was monitored. When an electric current through the etching capillary was observed, the capillary wall became porous. The etched part was fixed in a vial, where NaOH solution with a certain concentration was added during the sample injection. The whole capillary was filled with pH 3.0 running buffer. The inlet end vial was filled with protein sample dissolved in the running buffer. An electric voltage was applied across the inlet end vial and etched porous wall. A neutralization reaction occurs at the boundary (interface) of the fronts of H+ and OH-. A pH step or sharp pH gradient exists across the boundary. When positive protein ions electromigrate to the boundary from the sample vial, they are isoelectricelly focused at points corresponding to their pH. After a certain period of concentration, a high voltage is applied across the whole capillary and a conventional CE is followed. An over 100-fold concentration factor has been easily obtained for three model proteins (bovine serum albumin, lysozyme, ribonuclease A). Furthermore, the IEF sample concentration and its dynamics have been visually observed with the whole-column imaging technique. Its merits and remaining problem have been discussed, too.

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“…The CE instrument was similar to that used in our previous works [31,32]. A high-voltage power supply (Matsusada Precision, Osaka, Japan) provided injection and separation voltages.…”

Section: Capillary Electrophoresis Instrumentationmentioning

confidence: 99%

“…Etching of the capillary wall with HF has been reported [32,33]. In this work, we improved the HF etching method.…”

Section: Etching Of the Capillary Wallmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Isoelectric focusing sample injection for capillary electrophoresis of proteins

Wu¹,

Zhang²,

Onoda³

2005

Electrophoresis

Self Cite

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show abstract

“…13 A high-voltage power supply (Matsusada Precision Inc., Japan) provided injection and separation voltages. A UV absorbance detector (CE-970, Japan Spectroscopic Co. LTD) was used for indirect UV detection at 372 nm.…”

Section: Capillary Electrophoresis Instrumentationmentioning

confidence: 99%

Capillary Electrophoresis of Small Anions in Complex Samples with a Hollow Fiber Sampling Inlet End

Kasashima

Sakai

2003

ANAL. SCI.

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“…1,2 However, conventional CE is limited to a relatively poor concentration detection limit due to the low volume loadability of the capillary; 3,4 also, sample preconcentration is always considered before CE separation. [5][6][7][8][9][10][11][12][13][14][15] Among various concentration methods, isoelectric focusing (IEF) has been demonstrated to be an effective approach for the concentration of proteins. 14,15 In a conventional IEF, carrier ampholytes are used to establish a stable pH gradient.…”

Section: Introductionmentioning

confidence: 99%

An Improvement of the On-line Electrophoretic Concentration Method for Capillary Electrophoresis of Proteins and Experimental Factors Affecting the Concentration Effect

Cited by 19 publications

References 18 publications

Isoelectric focusing sample injection for capillary electrophoresis of proteins

Isoelectric focusing sample injection for capillary electrophoresis of proteins

Capillary Electrophoresis of Small Anions in Complex Samples with a Hollow Fiber Sampling Inlet End

Isoelectric Focusing Sample Injection for Capillary Zone Electrophoresis in a Fused Silica Capillary

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