Separation of basic proteins in free solution capillary electrophoresis: effect of additive, temperature and voltage

Cifuentes, Alejandro; Rodrı́guez, Miguel Á.; García-Montelongo, F.

doi:10.1016/0021-9673(96)00226-9

Cited by 74 publications

(50 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thus, gemini of 2 mM is selected as the optimal. Cifuentes et al [30] reported the separation of basic proteins using CTAB as the dynamic coating, and higher concentration (10 mM) of CTAB present in buffer and lower temperature (107C) were required to obtain an acceptable separation. Ding and Fritz [31] separated the basic proteins using a buffer containing less CTAB (0.6 mM), but the efficiencies of proteins were not reported.…”

Section: Protein Separationmentioning

confidence: 99%

Cationic gemini surfactant as dynamic coating in CE for the control of EOF and wall adsorption

Liu¹,

Li²,

Tang³

et al. 2007

Electrophoresis

View full text Add to dashboard Cite

The cationic gemini surfactant ethylene bis(1-dodecyldimethylammonium) dibromide was used as a dynamic coating to control EOF and prevent wall adsorption of basic proteins in CE for the first time. This gemini surfactant shows a more powerful capability in EOF reversal than traditional single-chained surfactant. The gemini surfactant reverses the EOF at a concentration level even less than 0.01 mM, and the EOF magnitude is affected by surfactant concentration, pH, ionic strength, and ions added in buffer. Highly efficient and rapid protein separation (N > 300,000) was obtained with buffer containing 2 mM gemini surfactant under pH ranging from 3 to 6. The effects of surfactant and buffer concentration on protein separation were investigated in detail. Under the optimal conditions, good repeatability (RSD of migration time <0.6% for run-to-run and <2.5% for day-to-day assays) and recovery (>90%) of tested proteins were obtained. This new dynamic coating is also suitable for biosample analysis.

show abstract

Section: Protein Separationmentioning

confidence: 99%

Cationic gemini surfactant as dynamic coating in CE for the control of EOF and wall adsorption

Liu¹,

Li²,

Tang³

et al. 2007

Electrophoresis

View full text Add to dashboard Cite

show abstract

“…Covalently linked or physically adsorbed polymer coating [7][8][9][10] provided good separation efficiency; however, it often suffered from short lifetime, troublesome preparation procedure, or unsatisfactory reproducibility. Dynamic coating [11,12] is a relatively attractive method due to its simplicity and versatility; however, the dynamic coating additive in buffer may interact with the analyte and ruin the separation or detection, e.g., it may interfere with ESI-MS detection. Lucy and co-workers [13][14][15] proposed a novel type of semipermanent capillary coatings based on the selfassembly of double-chained surfactants.…”

Section: Introductionmentioning

confidence: 99%

Long‐chained gemini surfactants for semipermanent wall coatings in capillary electrophoresis of proteins

Liu

Jie

et al. 2008

Electrophoresis

View full text Add to dashboard Cite

In this paper, we presented the first example of using gemini surfactants as semipermanent coatings in CE for protein separation. These coatings are based on the self-assembly of a series of cationic gemini surfactants, alkanediyl-alpha,omega-bis(dimethylalkylammonium bromide) (m-s-m), on the capillary wall. The coatings can keep stable for a long time without surfactant in the buffer, e.g., after the surfactants were removed from the buffer, the reversed EOF only decreased by 3.6 and 3.9% for 18-2-18 and 16-2-16 coatings over 60 min under continuous electrophoretic conditions. The coating stability increased with the alkyl chain length m. The double long chains of geminis (m > or = 14) yielded a good coating stability; meanwhile, the spacer group acted as an EOF modifier. Thus, this bifunctional surfactant coating provided a new buffer-independent method for EOF control. For 18-s-18 series, the best coating stability and largest EOF were obtained at s = 10. Ranging s from 3 to 10 yielded a linear fine-tuning of EOF and thereby allowed the adjustment of the protein apparent mobility. Highly efficient separation (>500 000 plates/m) was achieved with all the 18-s-18 coatings. Excellent run-to-run and day-to-day reproducibility (RSD of migration time

show abstract

“…These may be caused by the hydroxyl groups which possess higher hydrophilicity, and thus the HEC coating can be washed off the capillary wall easily due to its hydrophilicity. Therefore, in order to solve the insufficient adsorption of hydrophilic polymer on the capillary wall, a variety of positively charged polymers, such as polyethyleneimine [21,22]; a copolymer of vinylpyrrolidone and vinylimidazole [23]; Polybrene, poly(methoxyethoxyethyl) ethylenimine, poly(diallyldimethylammonium) chloride [24,25], ethylpyrrolidine methacrylate-N,N-dimethylacrylamide (EpyM-DMA) [26,27] were used as capillary coating for protein analysis by CE. Especially, cationic polysaccharides and their derivatives have attracted much attention for their both dynamic coating ability and reproducibility of separations [28][29][30][31][32][33].…”

Section: Introductionmentioning

confidence: 99%

Cationized hydroxyethylcellulose as a novel, adsorbed coating for basic protein separation by capillary electrophoresis

et al. 2008

View full text Add to dashboard Cite

We present cationized hydroxyethylcellulose (cat-HEC) synthesized in our laboratory as a novel physically adsorbed coating for CE. This capillary coating is simple and easy to obtain as it only requires flushing the capillary with polymer aqueous solution. A comparative study with and without polymers was performed. The adsorbed cat-HEC coating exhibited minimal interactions with basic proteins, providing efficient basic protein separations with excellent reproducibility. Under broad pHs, the amine groups are the main charged groups bringing about a global positive charge on the capillary wall. As a consequence, the cat-HEC coating produced an anodal EOF performance. A comparative study on the use of hydroxyethylcellulose (HEC) and cat-HEC as physically adsorbed coatings for CE are also presented. The separation efficiency and analysis reproducibility proved that the cat-HEC polymer was efficient in suppressing the adsorption of basic proteins onto the silica capillary wall. The long-term stability of the cat-HEC coating in consecutive protein separation runs has demonstrated the suitability of the coating for high-throughput electrophoretic protein separations.

show abstract

Separation of basic proteins in free solution capillary electrophoresis: effect of additive, temperature and voltage

Cited by 74 publications

References 23 publications

Cationic gemini surfactant as dynamic coating in CE for the control of EOF and wall adsorption

Cationic gemini surfactant as dynamic coating in CE for the control of EOF and wall adsorption

Long‐chained gemini surfactants for semipermanent wall coatings in capillary electrophoresis of proteins

Cationized hydroxyethylcellulose as a novel, adsorbed coating for basic protein separation by capillary electrophoresis

Contact Info

Product

Resources

About