Suppression of Electroosmotic Flow and Prevention of Wall Adsorption in Capillary Zone Electrophoresis Using Zwitterionic Surfactants

Yeung, Ken K.‐C.; Lucy, Charles A.

doi:10.1021/ac961231k

Cited by 121 publications

(139 citation statements)

References 32 publications

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“…This effect may be due to a reduction in dyedye quenching reactions on the protein surface due to denaturation of the protein in the presence of detergent, or possibly to the creation of hydrophobic environments around the attached dye molecules which promote fluorescence. Unlike SDS, the presence of DAPS does not alter the electrophoretic mobility of the proteins; however, the electroosmotic flow was reduced approximately twofold as has been reported previously [24].…”

Section: Cze Analysissupporting

confidence: 78%

Microchip separations of protein biotoxins using an integrated hand-held device

et al. 2005

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We report the development of a hand-held instrument capable of performing two simultaneous microchip separations (gel and zone electrophoresis), and demonstrate this instrument for the detection of protein biotoxins. Two orthogonal analysis methods are chosen over a single method in order to improve the probability of positive identification of the biotoxin in an unknown mixture. Separations are performed on a single fused-silica wafer containing two separation channels. The chip is housed in a microfluidic manifold that utilizes o-ring sealed fittings to enable facile and reproducible fluidic connection to the chip. Sample is introduced by syringe injection into a septum-sealed port on the device exterior that connects to a sample loop etched onto the chip. Detection of low nanomolar concentrations of fluorescamine-labeled proteins is achieved using a miniaturized laser-induced fluorescence detection module employing two diode lasers, one per separation channel. Independently controlled miniature high-voltage power supplies enable fully programmable electrokinetic sample injection and analysis. As a demonstration of the portability of this instrument, we evaluated its performance in a laboratory field test at the Defence Science and Technology Laboratory with a series of biotoxin variants. The two separation methods cleanly distinguish between members of a biotoxin test set. Analysis of naturally occurring variants of ricin and two closely related staphylococcal enterotoxins indicates the two methods can be used to readily identify ricin in its different forms and can discriminate between two enterotoxin isoforms.

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Section: Cze Analysissupporting

confidence: 78%

Microchip separations of protein biotoxins using an integrated hand-held device

et al. 2005

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“…As shown in Tab. 2, efficiency obtained for kin17 peak in our conditions is not as high as that reported previously for cationic proteins using permanent or dynamic coating [33,34]. This can be attributed mainly to the low ionic strength of the running buffer we employed, in order to avoid a dissociating effect of the BGE on the kin17-ssDNA complex formed.…”

Section: Analysis With a Peo-coated Capillarycontrasting

confidence: 48%

Poly(ethylene oxide) facilitates the characterization of an affinity between strongly basic proteins with DNA by affinity capillary electrophoresis

et al. 2005

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In order to study kin17 protein-DNA affinity, we have developed a fast and reproducible capillary electrophoresis (CE) analysis of a strongly basic protein: kin17 protein, using a nonpermanent coating based on poly(ethylene oxide) (PEO) to avoid adsorption of kin17. The coating procedure was optimized to provide a residual and stable electroosmotic flow (EOF = 5 x 10(-5) cm(2)/V x s), exhibiting RSD of 0.3% and excellent long-term stability. Good intraday and interday reproducibility of kin17 migration times (0.8 and 0.3% relative standard deviation (RSD), respectively) enabled us to consider that the recovery percentage obtained for kin17 protein was satisfactory (79%). The potential of this PEO-based coating procedure was evaluated for affinity CE method in order to study the affinity of kin17 protein for two single-stranded DNA (ssDNA) models: polydeoxyadenylic acid and polydeoxycytidilic acid (pdA and pdC). Binding constants (1.5 x 10(7) +/- 17% and 1.7 x 10(7) + 25%M(-1)) were evaluated assuming a 1:1 affinity between kin17 and pdA or pdC, respectively.

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“…Three basic proteins, ribonuclease A, cytochrome c and lysozyme were selected as model proteins due to their strong adsorption on the capillary wall, which results in bad efficiency, low protein recovery, and poor repeatability in migration time in common CE assay [13]. Generally speaking, an ideal coating for protein separation should exhibit high efficiency, good recovery and repeatability, retention of EOF, facility to apply, inexpensive, and availability over a wide range of buffer conditions [28,29]. Thus, we mainly focused on the performance of gemini dynamic coating in these aspects.…”

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

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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.

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Suppression of Electroosmotic Flow and Prevention of Wall Adsorption in Capillary Zone Electrophoresis Using Zwitterionic Surfactants

Cited by 121 publications

References 32 publications

Microchip separations of protein biotoxins using an integrated hand-held device

Microchip separations of protein biotoxins using an integrated hand-held device

Poly(ethylene oxide) facilitates the characterization of an affinity between strongly basic proteins with DNA by affinity capillary electrophoresis

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

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