High-Speed, Whole-Column Fluorescence Imaging Detection for Isoelectric Focusing on a Microchip Using an Organic Light Emitting Diode as Light Source

Yao, Bo; Yang, Huihua; Liang, Qionglin; Luo, Guoan; Wang, Liduo; Ren, Kangning; Gao, Yudi; Wang, Yiming; Qiu, Yong

doi:10.1021/ac060445r

Cited by 51 publications

(47 citation statements)

References 39 publications

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“…It has been performed in capillary format [8,9] or in microdevices [10][11][12][13][14][15], sometimes coupled with a second separation dimension [16][17][18][19]. Several IEF steps have even been coupled to improve protein resolution [20].…”

Section: Introductionmentioning

confidence: 99%

Use of quasi‐isoelectric buffers as anolyte and catholyte to improve capillary isoelectric focusing performances

2008

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The use of quasi-isoelectric anolytes and catholytes has been investigated to improve CIEF performances. Narrow pH cuts of carrier ampholytes (NC) have been compared to more conventional couples of anolytes/catholytes (phosphoric acid/sodium hydroxide and glutamic acid/lysine). First, a CIEF setup that consists in a bare silica capillary and 70:30 water/glycerol separation medium has been used. The experiments have shown that when using NC instead of more classical anolytes and catholytes, an increase in the protein detection time was observed and the resolutions obtained for neutral and acidic proteins were doubled. Moreover, according to the NC fraction used, the resolution was modified. In order to investigate further the mechanisms involved, a second setup using a capillary coated with hydroxypropylcellulose was used. With this setup no difference has been observed when changing anolyte and catholyte nature. A simple methodology has then been developed to evaluate EOF during focusing and mobilization steps of CIEF experiments. It highlighted the crucial role played by EOF when using a bare silica capillary. EOF indeed decreased by 33% during mobilization step when using NC instead of classical anolytes and catholytes.

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Section: Introductionmentioning

confidence: 99%

Use of quasi‐isoelectric buffers as anolyte and catholyte to improve capillary isoelectric focusing performances

2008

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“…To eliminate the mobilization step, whole-channel detection systems have been developed to reduce proteins' de-focusing during mobilization and improve separation resolution. The detection systems include a laser-induced fluorescence imaging system [27], organic light-emitting diode array [28] and a scanning system [29].…”

Section: Individual Dimension In Microfluidic Devicesmentioning

confidence: 99%

Two‐dimensional protein separation in microfluidic devices

Chen

Fan

2009

Electrophoresis

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Proteomics is emerging as an important tool in modern drug discoveries and medical diagnostics. One of the techniques used in proteomics studies is 2-DE. The process of the conventional 2-DE is time-consuming and it has substandard reproducibility. Many efforts have been made to address the limitations, with an aim for fast separation and high resolution. In this paper, we reviewed the work on achieving 2-DE in microfluidic devices, including individual dimension in one channel, two dimensions in two intersected channels, and 2-D separation in a large number of channels. We also discussed the need for integrating microvalves within 2-DE devices to prevent different separation media from contaminating with each other. Although more efforts are required to match the performance of conventional 2-DE in a slab gel, microfluidics-based 2-D separation has a potential to become an alternative in the future.

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“…The proteins stop their migration in the separation channel once they reach the pH zone corresponding to their pI. After reaching this equilibrium step, mobilization of the focused sample zones [91,92], which often requires additional instruments (e.g. micropumps or valves for the hydro- dynamic mode), is necessary to perform an in-channel detection at a single point of the separation channel.…”

Section: Cief On Chipsmentioning

confidence: 99%

“…To avoid mobilization step, whole-column imaging (WCI) method may be alternatively employed and was shown more ideal for on-chip CIEF. WCI detection is frequently carried out using an epifluorescence microscope with a xenon lamp [93] or a 100-W mercury lamp [79,94,95] or LED [92,[96][97][98]. Moreover, unstable EOF, pH gradient drift and compression of the pH gradient are significant problems leading often to poor resolution and reproducibility.…”

Section: Cief On Chipsmentioning

confidence: 99%

Recent innovations in protein separation on microchips by electrophoretic methods

Peng¹,

Pallandre²,

Tran³

et al. 2007

Electrophoresis

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Microchips for analytical purposes have attracted great attention over the last 20 years. In the present review, we focus on the most recent development of microchips for electrophoretic separation of proteins. This review starts with a short recalling about the microchips covering the basic microchip layout for CE and the commercial chips and microchip platforms. A short paragraph is dedicated to the surface treatment of microchips, which is of paramount importance in protein analysis. One section is dedicated to on-line sample pretreatment in microchips and summarizes different strategies to pre-concentrate or to purify proteins from complex matrixes. Most of the common modes used for CE of proteins have already been adapted to the chip format, while multidimensional approaches are still in progress. The different routes to achieve detection in microchip are also presented with a special attention to derivatization or labeling of proteins. Finally, several recent applications are mentioned. They highlight the great potential of electrophoretic separations of proteins in numerous fields such as biological, pharmaceutical or agricultural and food analysis. A bibliography with 151 references is provided covering papers published from 2000 to the early 2007.

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High-Speed, Whole-Column Fluorescence Imaging Detection for Isoelectric Focusing on a Microchip Using an Organic Light Emitting Diode as Light Source

Cited by 51 publications

References 39 publications

Use of quasi‐isoelectric buffers as anolyte and catholyte to improve capillary isoelectric focusing performances

Use of quasi‐isoelectric buffers as anolyte and catholyte to improve capillary isoelectric focusing performances

Two‐dimensional protein separation in microfluidic devices

Recent innovations in protein separation on microchips by electrophoretic methods

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