The effect of sample salt additives on capillary electrophoresis analysis of intact proteins using surface modified capillaries

Elhamili, Anisa; Wetterhall, Magnus; Puerta, Angel de la; Westerlund, Douglas; Bergquist, Jonas

doi:10.1016/j.chroma.2008.12.037

Cited by 7 publications

(4 citation statements)

References 38 publications

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“…This way, the use of high concentrations of ionic salt in BGE solutions has been one strategy for minimizing undesirable interactions between proteins and the surface of fused-silica capillaries. 38,40,41 On the other hand, this strategy must be carefully taken because this might lead to excessive Joule heating and/or denaturation of proteins. 42 The effect of the running buffer additive LiCl at 25 mmol L À1 on the IFN alpha-2a analysis was investigated.…”

Section: Methods Optimizationmentioning

confidence: 99%

Development, validation, and application of a capillary electrophoresis method for analysis of cytokine interferon alpha-2a in pharmaceutical formulations

2013

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A simple CE based method was developed and validated for the determination of interferon alpha-2a in a pharmaceutical formulation. After optimization, the best results were obtained using 30 mmol L À1 tetraborate buffer at pH 8.50 with 50 mmol L À1 of sodium dodecyl sulfate, and using a diode array detector at 200 nm. The applied voltage was +25 kV, and the sample injection was performed in the hydrodynamic mode. All analyses were carried out in a fused-silica uncoated capillary with an i.d. of 75 mm and an effective length of 50.0 cm. Under these conditions, the analysis was achieved in less than 12 min. Linearity was obtained in the range 0.41-1.54 MIU mL À1 (r $ 0.997). The RSD (%) and relative errors (%) obtained in precision and accuracy studies (intra-day and inter-day) were lower than 5%.Therefore, this method was found to be appropriate for controlling pharmaceutical formulations containing interferon alpha-2a.

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Section: Methods Optimizationmentioning

confidence: 99%

Development, validation, and application of a capillary electrophoresis method for analysis of cytokine interferon alpha-2a in pharmaceutical formulations

2013

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“…The effect of adding alkali salts to protein samples in the CE analysis of intact proteins was studied 91. A high degree of sample stacking even for large proteins was found to occur when alkali salts were added to the sample.…”

Section: Detection Schemes and Approachesmentioning

confidence: 99%

Electrophoretic and electrochromatographic separation of proteins in capillaries: An update covering 2007–2009

Rassi

2009

Electrophoresis

123

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This review article covers 3-year period from 2007 to 2009 and is a continuation of the review article by V. Dolnik, [Electrophoresis 2008, 29, 143-156]. This article with 125 references describes recent developments in CE and CEC of proteins in capillary format and does not cover the developments of CE and CEC in microchip format, since Tran et al. review the microchip subject in this special issue. The present review article has four major topics including (i) the separation media, (ii) multidimensional separations, (iii) detection, and (iv) applications.

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“…Transient isotachophoresis (ITP) − and dynamic pH junction − are often employed for capillary zone electrophoresis (CZE) of proteins. Using a mechanism similar to sweeping, protein peak height can be increased by adding anionic surfactants to the running buffer while sample is prepared without surfactants. , Protein peaks can be sharpened by adding salt or SDS , to the samples, generating a stacking effect. Insufficient surfactant can result in broad or multiple peaks because of heterogeneity of protein–surfactant complexes.…”

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confidence: 99%

Sample Stacking Provides Three Orders of Magnitude Sensitivity Enhancement in SDS Capillary Gel Electrophoresis of Adeno-Associated Virus Capsid Proteins

Zhang

Meagher

2017

Anal. Chem.

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Size-based protein analysis utilizing only 25 ng of total proteins has been realized by sodium dodecyl sulfate capillary gel electrophoresis (SDS CGE) with head-column field-amplified sample stacking as an online sample preconcentration technique. This method has been used as a replacement of SDS-PAGE for purity analysis of adeno-associated virus (AAV) therapeutic products of different serotypes and transgenes. A limit of detection of 0.2 ng/mL (3.3 pM) capsid proteins was achieved with convenient UV absorbance detection at 214 nm, equivalent to 20 pg of protein (330 attomole) loaded in the autosampler vial. For purity analysis, only 25 ng of total AAV capsid proteins (4.3 femtomole virus particles) were loaded to the autosampler vial. The sensitivity is comparable to silver-stained SDS-PAGE. The RSD of purity measurement was 0.0-0.8%, comparable to conventional SDS CGE utilizing 0.1-0.5 mg proteins. The new method provided 3 orders of magnitude sensitivity enhancement as compared to conventional SDS CGE. It shares all the advantages of conventional SDS CGE (labor-saving, easy automation, and convenient quantitation) and also the high sensitivity of silver stained SDS-PAGE. The sample stacking SDS CGE technique can be adopted for size-based analysis of other types of proteins. It is especially useful when protein quantity or concentration is not sufficient for regular SDS CGE or SDS-PAGE assay.

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The effect of sample salt additives on capillary electrophoresis analysis of intact proteins using surface modified capillaries

Cited by 7 publications

References 38 publications

Development, validation, and application of a capillary electrophoresis method for analysis of cytokine interferon alpha-2a in pharmaceutical formulations

Development, validation, and application of a capillary electrophoresis method for analysis of cytokine interferon alpha-2a in pharmaceutical formulations

Electrophoretic and electrochromatographic separation of proteins in capillaries: An update covering 2007–2009

Sample Stacking Provides Three Orders of Magnitude Sensitivity Enhancement in SDS Capillary Gel Electrophoresis of Adeno-Associated Virus Capsid Proteins

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