Increasing the Efficiency of In-Capillary Electrophoretically Mediated Microanalysis Reactions via Rapid Polarity Switching

Sanders, Brandi D.; Slotcavage, Rachel L.; Scheerbaum, Diana L.; Kochansky, Christopher J.; Strein, Timothy G.

doi:10.1021/ac048595u

Cited by 26 publications

(21 citation statements)

References 30 publications

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“…To improve the sensitivity and speed of EMMA methods, rapid polarity switching was introduced by Sanders et al. . It is based on the rapid (1–5 s) and sequential switching of the applied separation voltage polarity.…”

Section: Application Of On‐column Derivatisationmentioning

confidence: 99%

On‐capillary derivatisation as an approach to enhancing sensitivity in capillary electrophoresis

Glatz

2014

Electrophoresis

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Separation technologies play an important role in revealing biological processes at various omic levels, in pharmacological and clinical research. In this context, CE is a strong candidate for analyses of samples with rapidly increasing complexity. Even though CE is well known for its many advantages in this regard, the sensitivity of CE analyses is insufficient for many applications. Accordingly, there are generally three main options for enhancing the sensitivity of CE analyses - using special detection techniques, using sample pre-concentration and derivatisation. Derivatisation is often the method of choice for many laboratories, since it is simple and provides several advantages such as small sample volume demand and the possibility of automation. Although it can be performed in different ways depending on where the reaction takes place, this article reviews one of the simplest and at the same time most useful approaches on-capillary derivatisation. Even if in many cases the use of on-capillary derivatisation alone is enough to improve the detection sensitivity, on other occasions it needs to be employed in combination with the other above-mentioned strategies. After a simple discussion of derivatisation in general, special attention is focused on the on-capillary approach and methodologies available for on-capillary reactant mixing. Its applications in various fields are also described.

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Section: Application Of On‐column Derivatisationmentioning

confidence: 99%

On‐capillary derivatisation as an approach to enhancing sensitivity in capillary electrophoresis

Glatz

2014

Electrophoresis

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“…The first is to apply a series of positive and negative voltages across the capillary. This rapid polarity switching improves the convection of reactant plugs due to forward and backward shaking movements. The second approach is to combine the effect of the electric field (EMMA) with an amplification step at zero potential (zero‐potential amplification) .…”

Section: In‐capillary Assaysmentioning

confidence: 99%

Advances in capillary electrophoresis for miniaturizing assays on kinase enzymes for drug discovery

Nehmé

Morin

2015

Electrophoresis

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CE has become a frequently used tool for miniaturizing enzyme assays due particularly to its well-recognized low sample consumption. CE-based enzyme assays cover all aspects of kinetic analysis including the evaluation of enzyme activity, substrate and modulator characterization, and identification. These assays are performed to conduct high-quality primary (hit finding) and secondary (confirmation by determining the half maximal inhibitory concentration, IC ) screening. Nowadays, the kinase family is among the most actively studied pharmaceutical targets in oncology, and in neurodegenerative and inflammation diseases. In this article, we review the fundamentals of the different approaches that may be employed for assaying kinase kinetics. Their advantages and limitations will also be discussed by covering the literature of CE-based assays for purified kinases as well as for kinases in living cells. The last section will be devoted to perspectives by showing some applications of multiplexed and of microchip devices for high-throughput screening kinase assays.

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“…A zero-potential amplification is possible whereby the voltage is stopped when the two zones have merged, to allow accumulation of more product, and therefore to increase the sensitivity. Recently, Strein and co-workers [32] introduced a new approach to enhance the sensitivity and speed of EMMA with small molecules. Rapid polarity switching (RPS) is used as a novel technique that takes advantage of the differences in electrophoretic mobilities of two reagents to produce successive mixing events through repetitive reversals of the applied potential.…”

Section: Zone-passing Techniquementioning

confidence: 99%

Advances in capillary electrophoretically mediated microanalysis: An update

2006

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This review, as a continuation of an earlier report, gives an overview of recent developments, over the period from 2003 until now, in the use of capillary electrophoretic techniques for the in-line study of enzymatic reactions, derivatization, and chemical reactions. The article is divided into two parts: (i) in-line enzymatic reactions and (ii) in-line derivatization and chemical reactions. The first part introduces electrophoretically mediated microanalysis (EMMA) and discusses and illustrates the different modes of EMMA. A literature overview on enzymatic reactions is provided. The second part starts with an introduction of the procedures and the nomenclature used in the area of in-line derivatization and chemical reactions based on EMMA. Reported derivatization and chemical reaction procedures are discussed and summarized.

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Increasing the Efficiency of In-Capillary Electrophoretically Mediated Microanalysis Reactions via Rapid Polarity Switching

Cited by 26 publications

References 30 publications

On‐capillary derivatisation as an approach to enhancing sensitivity in capillary electrophoresis

On‐capillary derivatisation as an approach to enhancing sensitivity in capillary electrophoresis

Advances in capillary electrophoresis for miniaturizing assays on kinase enzymes for drug discovery

Advances in capillary electrophoretically mediated microanalysis: An update

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