Capillary zone electrophoresis for the determination of electrophoretic mobilities and diffusion coefficients of proteins

Walbroehl, Yvonne; Jorgenson, James W.

doi:10.1002/mcs.1220010110

Cited by 108 publications

(43 citation statements)

References 18 publications

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“…In addition, many physical properties of proteins can be measured by CE usually requiring other sophisticated equipment. Notable among these are thermodynamic constants [20], melting temperatures [19,20], diffusion coefficients [21], and "native" (vs. denatured) pIs [17].…”

Section: Principlesmentioning

confidence: 99%

Capillary electrophoresis and the clinical laboratory

et al. 2006

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Over the past 15 years, CE as an analytical tool has shown great promise in replacing many conventional clinical laboratory methods, such as electrophoresis and HPLC. CE's appeal was that it was fast, used very small amounts of sample and reagents, was extremely versatile, and was able to separate large and small analytes, whether neutral or charged. Because of this versatility, numerous methods have been developed for analytes that are of clinical interest. Other than molecular diagnostic and forensic laboratories CE has not been able to make a major impact in the United States. In contrast, in Europe and Japan an increasing number of clinical laboratories are using CE. Now that automated multicapillary instruments are commercially available along with cost-effective test kits, CE may yet be accepted as an instrument that will be routinely used in the clinical laboratories. This review will focus on areas where CE has the potential to have the greatest impact on the clinical laboratory. These include analyses of proteins found in serum and urine, hemoglobin (A1c and variants), carbohydrate-deficient transferrin, forensic and therapeutic drug screening, and molecular diagnostics.

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Section: Principlesmentioning

confidence: 99%

Capillary electrophoresis and the clinical laboratory

et al. 2006

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“…Many different techniques have been developed for reducing the adsorption of proteins to the walls of the channels. Some examples include the use of pH extremes, high-concentration buffers, and surface coatings that are tailored for both the surface chemistry of the substrate and the proteins [2][3][4][5][6][7][8][9]. Most of these methods have limitations that reduce their overall applicability.…”

Section: Introductionmentioning

confidence: 99%

Micellar electrokinetic chromatography of fluorescently labeled proteins on poly(dimethylsiloxane)‐based microchips

et al. 2006

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MEKC of standard proteins was investigated on PDMS microfluidic devices. Standard proteins were labeled with AlexaFluor(R) 488 carboxylic acid tetrafluorophenyl ester and filtered through a size-exclusion column to remove any small peptides and unreacted label. High-efficiency MEKC separations of these standard proteins were performed using a buffer consisting of 10 mM sodium tetraborate, 25 mM SDS, and 20% v/v ACN. A separation of BSA using this buffer in a 3.0 cm long channel generated a peak with a plate height of 0.38 microm in <20 s. Additional fast separations of myoglobin, alpha-lactalbumin, lysozyme, and cytochrome c also yielded peaks with plate heights ranging from 0.54 to 0.72 microm. All proteins migrated with respect to their individual pIs. To improve the separations, we used a PDMS serpentine chip with tapered turns and a separation distance of 25 cm. The number of plates generated increased linearly with increasing separation distance on the extended separation channel chips; however, the resolution reached an asymptotic value after about 7 cm. This limited the peak capacity of the separation technique to 10-12.

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“…The diffusion coefficients of β-lactoglobulin A and B were determined in triplicates by FFT method in three time intervals (Table S- 4). The collected data became more scattered as the measurement time was increased.…”

Section: Accurate Determination Of Diffusion Coefficientmentioning

confidence: 99%