Separation of β-lactoglobulin A, B and C variants of bovine whey using capillary zone electrophoresis

Paterson, Geoff R.; Hill, J. P.; Otter, Don

doi:10.1016/0021-9673(95)00054-q

Cited by 33 publications

(8 citation statements)

References 8 publications

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“…EOF decreases by two orders of magnitude when the microchannel was filled with 0.5% MC. Figure 4 shows that the EOF in the PMMA microchannel increases by adding Tween 20 which is an EOF suppressor in fused-silica CE [25][26][27]. Also, because of the high EOF mobility, the electrophoresis was not successful in the Tris-HCl buffer which includes Tween 20.…”

Section: Eof Measurementsmentioning

confidence: 93%

Dynamic coating using methylcellulose and polysorbate 20 for nondenaturing electrophoresis of proteins on plastic microchips

Mohamadi¹,

Mahmoudian²,

Kaji³

et al. 2007

Electrophoresis

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A dynamic coating using methylcellulose (MC) and a nonionic detergent (polysorbate 20) was developed, which controlled protein adsorption onto the surface of microchannels on a microchip made of poly(methyl methacrylate) (PMMA). Optimum concentration of polysorbate 20 in combination with the range of MC concentrations controlled the protein adsorption onto the microchannel surface, and increased the solubility of the protein samples while facilitating the injection of high concentrations of MC solutions into the microchannels. Higher concentrations of nonionic detergent increased the EOF mobility as opposed to the electrophoretic mobility and caused the electrophoresis to fail. Nondenaturing microchip electrophoresis of protein samples with molecular masses ranging from 20 to 100 kDa were completed in 100 s. Also, successful separation of a BSA sample and its complex with anti-BSA mAb ( 220 kDa) was achieved on a PMMA microchip. The separation exhibited high reproducibility in both migration time (RSD = 1%) and peak area (RSD = 10-15%).

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Section: Eof Measurementsmentioning

confidence: 93%

Dynamic coating using methylcellulose and polysorbate 20 for nondenaturing electrophoresis of proteins on plastic microchips

Mohamadi¹,

Mahmoudian²,

Kaji³

et al. 2007

Electrophoresis

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“…This class comprises a vast number of compounds, starting from monoamines, such as triethylamine and propylamine [24,25], morpholine [26], glucosamine and galactosamine [27], N,N-diethylethanolamine, N-ethyldiethanolamine, triethanolamine [28], ethanolamine [29], hydroxylamine, ethylamine [30], as well as the quaternary base tetramethylammonium chloride [31]. Among the diamines: 1,3-diaminopropane [32], 1,4-diaminobutane (putrescine) [33], 1,5-diaminopentane (cadaverine) [34], ethylenediamine [35], N,N,N©,N©-tetramethyl-1,3-butanedia- [36], and the a,w-bis-quaternary ammonium alkanes, such as hexamethonium [37] and decamethonium bromides [38].…”

Section: Amino Quenchersmentioning

confidence: 99%

The state of the art of dynamic coatings

et al. 2001

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The present review highlights the mechanisms of action and efficiency of three major classes of dynamic coatings so far adopted in capillary electrophoresis: (i) amines to oligo-amines, (ii) neutral synthetic and natural polymers, and (iii) neutral and zwitter-ionic surfactants. Their merits and efficacy have been explored in depth via a novel quantitation technique consisting of eluting, by frontal analysis, any adsorbed proteinaceous material, which can then be correctly quantified as a peak as it moves in front of the detector window. This is achieved by loading sodium dodecyl sulfate (SDS) micelles onto the cathodic side and migrating them electrophoretically into the capillary lumen, where they efficiently sweep any adsorbed polypeptide material. It is found that a common trend, for all quenchers, is linked to a hydrophobicity scale: the more hydrophobic the inhibitor, the better it minimizes potential interactions of macromolecules with the wall. This seems to be true for all the classes of dynamic modifiers tested. Finally, we describe a novel, dynamic to static quencher: it is a quaternary piperazine, bearing a reactive iodine atom at the end of a butyl tail (N(methyl-N-omega-iodo-butyl),N'-methyl piperazine). This molecule first binds to the wall, at alkaline pH values, via ionic and hydrogen bonds. Once docked onto the wall, the reactive tail forms a covalent link with the silica surface, to which it then remains permanently affixed.

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“…At high pH (i.e., pH 4 7), both the whey proteins and the capillary wall are negatively charged, and the adsorption process is minimized as a result of a charge repulsion effect. Good examples of this strategy are the separations obtained by Recio et al [26], Paterson et al [27], Paterson et al [28] and Kinghorn et al [29]. In all these methods, uncoated capillaries were employed and the adsorption of proteins to the capillary wall was reduced by the combination of high pH and high-ionic strength separation buffers with added alkali metal salts [26] or nonionic surfactants (e.g., Tween 20) [27][28][29].…”

Section: Separation Of Bovine Milk Proteinsmentioning

confidence: 99%

“…At high pH, separation of the genetic variants of b-Lg is possible, because the A variant has one negative charge more than the B variant (Asp 64 /Gly 64 ), and therefore, displays a slower migration towards the cathode. The genetic variant C of b-Lg was also separated using a high pH buffer, specifically 50 mM MES at pH 8.0, with the addition of 0.1% Tween 20 [27,28]. b-Lg C presents a higher positive charge than the A and B variants (Gln 59 of variant B is substituted by His 59 in variant C), and, therefore, exhibits the shortest migration time.…”

Section: Analyses Of Milk From Species Other Than Bovine and Genetic mentioning

confidence: 99%

Capillary electrophoresis for the analysis of food proteins of animal origin

2001

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In recent years, capillary electrophoresis (CE) has become an analytical technique with many applications in the study of food proteins and peptides. This review describes the existing CE methods of analysis of milk, egg, meat and fish proteins and peptides. The major developments in the application of CE to solve different problems in food technology, such as the assessment of technological processes, quality, and authenticity control of animal foods, are considered. A section dealing with future directions on the analysis of food proteins by CE is also included.

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Separation of β-lactoglobulin A, B and C variants of bovine whey using capillary zone electrophoresis

Cited by 33 publications

References 8 publications

Dynamic coating using methylcellulose and polysorbate 20 for nondenaturing electrophoresis of proteins on plastic microchips

Dynamic coating using methylcellulose and polysorbate 20 for nondenaturing electrophoresis of proteins on plastic microchips

The state of the art of dynamic coatings

Capillary electrophoresis for the analysis of food proteins of animal origin

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