Adsorption of proteins to fused-silica capillaries as probed by atomic force microscopy

Bonvent, Jean-Jacques; Barberi, R.; Bartolino, Roberto; Capelli, Laura; Righetti, Pier Giorgio

doi:10.1016/s0021-9673(96)00631-0

Cited by 37 publications

(26 citation statements)

References 28 publications

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“…On the other hand, it cannot be excluded that this lack of migration time reproducibility could be due, among other factors, to protein adsorption even if coated capillaries are used [35]. This adsorption to the capillary wall happens in a protein concentration-dependent manner [36], so that samples with high AGP concentration might affect reproducibility in a larger extent than samples with lower AGP concentration.…”

Section: Intraday Migration Time Precisionmentioning

confidence: 99%

CIEF with hydrodynamic and chemical mobilization for the separation of forms of α‐1‐acid glycoprotein

Lacunza¹,

Diez-Masa²,

Frutos³

2007

Electrophoresis

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Abstractα‐1‐Acid glycoprotein (AGP) is a protein that exists in different forms, which is due to variations in the amino acid sequence and/or in the glycosidic part of the protein. These differences confer to these forms, among other characteristics, diverse pIs. Changes in these forms of AGP have been correlated to modifications of the pathophysiological conditions of the individuals. One of the analytical techniques employed for their study has been IEF performed in slab gels. CIEF method with hydrodynamic and chemical mobilization, involving an isotachophoretic process, is developed in this work to separate up to 12 bands of forms of standard AGP, which is proposed as a more reproducible, quantitative, less sample‐consuming, and more automated one than conventional IEF. The challenge of this work has been the development of a CIEF method for the separation of bands of a very acidic protein (pI range: 1.8–3.8) in a capillary. Intraday RSD values ≤ 1.7% have been achieved for the relative migration time of the AGP bands to that of an internal standard. For intraday area precision, RSD (%) in the range of 2.70–22.71% for AGP zones accounting for more than 10% of total area of AGP sample has been obtained. As a proof of the potential of the methodology proposed, an AGP sample purified from a pool of sera of patients suffering from ovary cancer is analyzed by CIEF.

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Section: Intraday Migration Time Precisionmentioning

confidence: 99%

CIEF with hydrodynamic and chemical mobilization for the separation of forms of α‐1‐acid glycoprotein

Lacunza¹,

Diez-Masa²,

Frutos³

2007

Electrophoresis

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“…Even at such low pH values, adsorption still occurred and could only be prevented by addition of short-chain hydroxyethyl cellulose, which formed a dynamic coating. In our case we further checked protein recovery by frontal analysis, i.e., injecting SDS micelles which would desorb any adsorbed proteinaceous material [41]. Although we seem to have all the correct indicators in order, it is clear from our data that reproduction in transit times cannot be obtained easily.…”

Section: Facts and Misfacts On Cze Of Proteinsmentioning

confidence: 92%

Variety identification in maize linesvia capillary electrophoresis of zeins in isoelectric acidic buffers

et al. 1999

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“…It is well known that proteins tend to adsorb onto the capillary walls; indeed, this has recently been proven by direct probing of the surface using AFM [81]. DNA molecules tend not to adsorb on the silica surface due to electrostatic repulsion between the negatively charged wall and the phosphodiester backbone of DNA.…”

Section: Channel Coating and Polymer Solutionmentioning

confidence: 99%

DNA sequencing by capillary array electrophoresis and microfabricated array systems

Carrilho

2000

Electrophoresis

113

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To comply with the current needs for high‐speed DNA sequencing analysis, several instruments and innovative technologies have been introduced by several groups in recent years. This review article discusses and compares the issues regarding high‐throughput DNA sequencing by electrophoretic methods in miniaturized systems, such as capillaries, capillary arrays, and microchannels. Initially, general features of several capillary array designs (including commercial ones) will be considered, followed by similar analyses with microfabricated array electrophoretic devices and how they can contribute to the success of large sequencing projects.

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Adsorption of proteins to fused-silica capillaries as probed by atomic force microscopy

Cited by 37 publications

References 28 publications

CIEF with hydrodynamic and chemical mobilization for the separation of forms of α‐1‐acid glycoprotein

CIEF with hydrodynamic and chemical mobilization for the separation of forms of α‐1‐acid glycoprotein

Variety identification in maize linesvia capillary electrophoresis of zeins in isoelectric acidic buffers

DNA sequencing by capillary array electrophoresis and microfabricated array systems

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