Utility of high resolution capillary electrophoresis for monitoring peptide homo- and hetero-dimer formation

Landers, James P.; Oda, Robert P.; Liebenow, Jane A.; Spelsberg, Thomas C.

doi:10.1016/0021-9673(93)80651-n

Cited by 17 publications

(5 citation statements)

References 8 publications

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“…Boric acid is able to complex with diol groups, which can facilitate the analysis of sugars and glycoproteins (Landers et al 1993). It has been successfully utilized for the separation of another glycoprotein, IgG, in bovine colostrum products in our laboratory (Zhao et al 2007).…”

Section: Optimization Of Ce Conditionsmentioning

confidence: 99%

Analysis of α-lactalbumin, β-lactoglobulin A and B in whey protein powder, colostrum, raw milk, and infant formula by CE and LC

Ding

Yang

Zhao

et al. 2011

Dairy Science & Technol.

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Two analytical methods, capillary electrophoresis (CE) with UV detection and high-performance liquid chromatography (HPLC) with photodiode array detection, were developed for the determination of α-lactalbumin (α-Lac), β-lactoglobulin A (β-Lg A), and β-lactoglobulin B (β-Lg B) in whey protein powder, colostrum, raw milk, and infant formula. The CE analysis was performed in a bare fused silica capillary (57 cm×50 μm, effective length 50 cm) with a separation buffer consisting of 0.5 mol L −1 boric acid, 0.025 mol L −1 hydroxypropyl-β-cyclodextrin, and 0.8 g L −1 poly(ethylene oxide) 4,000,000 at pH 9.10. The HPLC analysis was performed on a CAPCELL PAK C 8 SG 300 (200×4.6 mm, 5-μm particles) column with a gradient mobile phase mixture consisting of acetonitrile and water containing trifluoroacetic acid. The CE method is suitable for the simultaneous determination of α-Lac, β-Lg A, and β-Lg B in whey powder, colostrum, and raw bovine milk, whereas the HPLC method can only be used for quantitative assays of β-Lg A and β-Lg B due to the co-elution of lactoferrin with α-Lac. Recoveries for both methods over the concentration range were between 90.7% and 116.8%. Whey protein powder had the highest content of the above three proteins. The contents of α-Lac in infant formula increased with the raise of age stage. Infant formula with the same age stage but different brands and countries had significant differences in the content of α-Lac.

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Section: Optimization Of Ce Conditionsmentioning

confidence: 99%

Analysis of α-lactalbumin, β-lactoglobulin A and B in whey protein powder, colostrum, raw milk, and infant formula by CE and LC

Ding

Yang

Zhao

et al. 2011

Dairy Science & Technol.

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“…It has been shown that CE can provide information on identity, purity, and structural changes of peptides [8][9][10][11]. In recent years, CE-MS coupling has become an important if not indispensable tool in this field of research [12][13][14][15][16][17][18][19][20].…”

Section: Introductionmentioning

confidence: 99%

Capillary electrophoresis‐mass spectrometry of peptides from enzymatic protein hydrolysis: Simulation and optimization

Simó

Cifuentes

2003

Electrophoresis

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Two important limitations still exist for the characterization of protein digests by capillary electrophoresis-mass spectrometry (CE-MS): (i) the buffer choice (i.e., the buffer must provide an adequate CE separation without ruining the MS signal), and (ii) the frequent generation of "unexpected" peptidic fragments during the enzymatic protein hydrolysis. In this work, a new approach is used to solve these difficulties, namely a theoretical model that relates the electrophoretic behavior of peptides to their sequence. The effectiveness of this procedure is demonstrated by the fast attainment of good CE-MS conditions for analyzing the peptides obtained from an enzymatic protein hydrolysate in a single run. This strategy can provide useful information for helping to characterize "unexpected" fragments from protein digests.

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“…Separation of peptides by capillary electrophoresis (CE) has become one of the most important application fields of this technique [1][2][3][4][5][6]. By using CE it has been possible to obtain valuable information on identity, purity and structural changes of peptides [7][8][9][10]. Moreover, the high efficiencies usually obtained (in the range of hundreds of thousands of plates) together with the short analysis times (lower than 30 min) and the compatibility with small sample volumes have made of CE a major laboratory tool for the separation, analysis and characterization of such biomolecules [11,12].…”

Section: Introductionmentioning

confidence: 99%

“…A second goal, when real samples are studied, is to fully identify the peptides. This can be done by matching peak position with standards [10] or if they are not available by techniques such as HPLC-MS [13,14,16], UV scanning [17], laser-induced fluorescence with selective derivatizing reagents [12], radioactive labeling [18], etc. In recent years, CE-MS has become an important if not indispensable tool in this field of research [19][20][21][22][23][24][25][26][27].…”

Section: Introductionmentioning

confidence: 99%

Simulation and optimization of peptide separation by capillary electrophoresis-mass spectrometry

2002

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The potential of capillary electrophoresis (CE) for the separation of peptides has been extensively demonstrated in the last decade. Their correct characterization and sequenciation is a difficult task that can be accomplished using CE-mass spectrometry (CE-MS). An important limitation of CE-MS is the buffer choice since it should provide an adequate CE separation without ruining the MS signal. In this work, a new strategy is used to help to solve this limitation based on the combination of two different methodologies. Namely, an ab initio semiempirical model that relates electrophoretic behavior of peptides to their sequence is first used to obtain in a fast and easy way adequate CE buffers compatible with MS analysis. Next, CE-MS is used to separate and characterize peptides via the determination of their relative molecular masses. The usefulness of this procedure is demonstrated analyzing in a single CE-MS run a group of 10 standard peptides of very different nature (i.e., relative molecular masses ranging from 132 to 1037 and isoelectric points ranging from 5.69 to 10.62). It is concluded that the use of this strategy can help to overcome the buffer limitation in CE-MS.

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Utility of high resolution capillary electrophoresis for monitoring peptide homo- and hetero-dimer formation

Cited by 17 publications

References 8 publications

Analysis of α-lactalbumin, β-lactoglobulin A and B in whey protein powder, colostrum, raw milk, and infant formula by CE and LC

Analysis of α-lactalbumin, β-lactoglobulin A and B in whey protein powder, colostrum, raw milk, and infant formula by CE and LC

Capillary electrophoresis‐mass spectrometry of peptides from enzymatic protein hydrolysis: Simulation and optimization

Simulation and optimization of peptide separation by capillary electrophoresis-mass spectrometry

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