Adaptation of Capillary Isoelectric Focusing to Microchannels on a Glass Chip

Hofmann, Oliver; Che, Diping; Cruickshank, Kenneth A.; Müller, Uwe R.

doi:10.1021/ac9806660

Cited by 165 publications

(140 citation statements)

References 21 publications

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“…The method offers high efficiency, versatility, speed, and economy of analysis (i.e., very low consumption of reagents and analytes; ref. [6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22]. Considerable effort also has been directed toward miniaturization in the field of CE-primarily in the adaptation of CE from capillaries to a planar microchannel format (9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19).…”

mentioning

confidence: 99%

SDS capillary gel electrophoresis of proteins in microfabricated channels

Yao

Anex

Caldwell

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

130

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Analysis of variations in the concentrations or structures of biomolecules (e.g., mRNAs, proteins, peptides, natural products) that occur either naturally or in response to environmental or genetic perturbations can provide important insight into complex biological processes. Many biological samples are mixtures that require a separation step before quantitation of variations in the individual components. Twodimensional denaturing gel electrophoresis has been used very effectively to separate complex mixtures of proteins, but it is time consuming and requires considerable amounts of sample. Microchannel-based separations have proven very effective in rapidly separating small amounts of nucleic acids; more recently, isoelectric focusing of proteins also has been adapted to the microchannel format. Here, we describe microchannel-based SDS capillary gel electrophoresis of proteins and demonstrate the speed and high resolution it provides. This development is an important step toward the miniaturization and integration of multidimensional and array separation methods for complex protein mixtures.

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mentioning

confidence: 99%

SDS capillary gel electrophoresis of proteins in microfabricated channels

Yao

Anex

Caldwell

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

130

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show abstract

“…Many researchers are developing completely integrated systems or lab-on-a-chip devices for biochemical analysis that accept a sample, perform a multistep process, and analyze the result (Service 1998a,c;Kricka 1998;Hofmann et al 1999). These steps may include extraction, purification, fluid movement, fluid mixing, reactions, and analysis.…”

Section: Mass Spectrometrymentioning

confidence: 99%

Automation for Genomics, Part Two: Sequencers, Microarrays, and Future Trends

Meldrum¹

2000

Genome Res.

116

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Automation for genomics has enabled a 43-fold increase in the total finished human genomic sequence in the world in the past four years. This is the second half of a two-part, noncomprehensive review that presents an overview of different types of automation equipment used in genome sequencing. The first part of the review, published in the previous issue, focused on automated procedures used to prepare DNA for sequencing or analysis. This second part of the review presents a look at available DNA sequencers and array technology and concludes with a look at future technologies. Alternate sequencing technologies including mass spectrometry, biochips, and single molecule analysis are included in this review.Sequencing data, essential for many of the medical breakthroughs of today and tomorrow, is currently accumulating at an exponential rate, largely because of advances in sequencing methods and laboratory automation. Instruments are available that can automate nearly every step in the large-scale sequencing process.The first article in this two-part review (Meldrum 2000) presented a description of automation designed for isolating DNA, cloning or amplifying DNA, preparing enzymatic sequencing reactions, and purifying DNA. Part one of the review also showed that the majority of genome centers use both in-house automation and commercial automation (Collins et al. 1998;Wendl et al. 1998;Mullikin and McMurragy 1999;Spurr et al. 1999; see http://www.genome.org for a supplementary table providing a snapshot view of automation used at a number of different genome centers).In this, the second part of the review, the focus is on automation after DNA preparation and sequencing reactions-on obtaining the sequence and its analysis. A discussion of future technologies for DNA sequencing projects is also included. (See the web sites referenced for photos or further details on the instruments presented.)

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“…In conventional CIEF, however, the focused analytes must be transported toward a detection window, which can sometimes decrease the resolution due to a band broadening during transportation [5]. Recently, many IEF techniques using microfluidic devices (microfluidic IEF, MIEF) have been developed to overcome this drawback [6][7][8][9][10][11][12]. MIEF has many advantages compared to CIEF, including a short analysis time (within 5 min) due to a short separation channel (several cm) and the requirement of a minimal amount of reagents and samples (several L).…”

Section: Introductionmentioning

confidence: 99%

A Simple and Easy-to-Use Capillary Isoelectric Focusing Technique Using Reagent-Release Hydrogels

et al. 2017

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Reagent-release hydrogels containing an acid or a base solution were prepared and employed as ion suppliers in isoelectric focusing using short capillaries. In conventional isoelectric focusing techniques using microfluidic channels, complicated liquid manipulations are required, which can decrease analytical performance such as the reproducibility of a focusing position. To develop a rapid and sensitive assay based on microfluidic isoelectric focusing, we previously prepared reagent-release capillaries retaining ampholytes by physical adsorption on the inner surface of a short capillary, which were then applied to isoelectric focusing. In this study, reagent-release hydrogels containing an acid or a base solution were developed for isoelectric focusing using short capillaries in place of the acid and base solutions used in conventional microfluidic isoelectric focusing. The prepared reagent-release hydrogels showed some advantages for isoelectric focusing, e.g., high durability for at least five weeks of storage, easy handling as compared to the conventional solutions, and suppression of hydrodynamic flow in capillaries, which often decreases the reproducibility in capillary isoelectric focusing. The migration behavior of ions from the hydrogels into the capillary and the formation of a pH gradient in isoelectric focusing using the reagent-release hydrogels were investigated, which confirmed that the prepared reagent-release hydrogels could form a similar pH gradient to that obtained with conventional capillary isoelectric focusing using the solutions.

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Adaptation of Capillary Isoelectric Focusing to Microchannels on a Glass Chip

Cited by 165 publications

References 21 publications

SDS capillary gel electrophoresis of proteins in microfabricated channels

SDS capillary gel electrophoresis of proteins in microfabricated channels

Automation for Genomics, Part Two: Sequencers, Microarrays, and Future Trends

A Simple and Easy-to-Use Capillary Isoelectric Focusing Technique Using Reagent-Release Hydrogels

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