Capillary Electrophoresis

Monnig, Curtis A.; Kennedy, Robert T.

doi:10.1021/ac00084a013

Cited by 296 publications

(88 citation statements)

References 452 publications

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“…[1][2][3][4] for exhaustive and up-to-date reviews). The most successful methods are the ones whereby the silanol groups are shielded by a polymer layer [5][6][7][8][9][10][11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

Performance of a physically adsorbed high-molecular-mass polyethyleneimine layer as coating for the separation of basic proteins and peptides by capillary electrophoresis

Erim

Cifuentes

Poppe

et al. 1995

Journal of Chromatography A

154

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Performance of a physically adsorbed high-molecular-mass polyethyleneimine layer as coating for the separation of basic proteins and peptides by capillary electrophoresis Erim, F.B.; Cifuentes, A.; Poppe, H.; Kraak, J.C. General rightsIt is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), other than for strictly personal, individual use, unless the work is under an open content license (like Creative Commons). Disclaimer/Complaints regulationsIf you believe that digital publication of certain material infringes any of your rights or (privacy) interests, please let the Library know, stating your reasons. In case of a legitimate complaint, the Library will make the material inaccessible and/or remove it from the website. Please Ask the Library: http://uba.uva.nl/en/contact, or a letter to: Library of the University of Amsterdam, Secretariat, Singel 425, 1012 WP Amsterdam, The Netherlands. You will be contacted as soon as possible. AbstractA simple method for the preparation of a polyethyleneimine (PEI) coating on the inner surface of fused-silica capillaries for capillary electrophoresis (CE) is reported. The PEI layer can be coated on the silica surface by just flushing the capillary with a solution containing high-molecular-mass PEI. The physically adsorbed layer appears to be very stable and can be used in a pH range of 3-11. In comparison to described methods to fabricate an immobilized PEI layer, the proposed method does not require an immobilization step, is simple and the preparation time of the coating is less than two hours. Good reproducibilities of migration times of basic proteins and peptides were obtained on the same PEI-coated capillary as well as on different PEI-coated capillaries. For basic proteins efficiencies ranging from 300 000-500 000 plates per meter were normally found.

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Section: Introductionmentioning

confidence: 99%

Performance of a physically adsorbed high-molecular-mass polyethyleneimine layer as coating for the separation of basic proteins and peptides by capillary electrophoresis

Erim

Cifuentes

Poppe

et al. 1995

Journal of Chromatography A

154

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“…Capillary electrophoresis (CE), 1 initially performed by Jorgenson and Lukacs (2), has emerged in recent years as a versatile and powerful tool for the analysis of biological molecules, because of its unparalleled efficiency for separation, small injection volumes, and low mass detection limits (3,4). We recently developed a CE method for the separation of peptides cleaved from hydrophobic membrane proteins, such as bacteriorhodopsin, a possible model for the study of G protein-coupled receptors (5).…”

mentioning

confidence: 99%

Structurally Related Peptide Agonist, Partial Agonist, and Antagonist Occupy a Similar Binding Pocket within the Cholecystokinin Receptor

Dong

Ding

Pinon

et al. 1999

Journal of Biological Chemistry

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The molecular basis of ligand binding to receptors provides important insights for drug development. Here, we explore domains of the cholecystokinin (CCK) receptor that are critical for ligand binding, using a novel series of fluorescent photolabile probes, receptor proteolysis, and rapid high resolution separation of peptide fragments by capillary electrophoresis. Each probe incorporated the same fluorophore and a photolabile p-benzoylphenylalanine at the amino terminus of the pharmacophoric domain (residue 24 of CCK-33) of CCK analogues representing full agonist, partial agonist, and antagonist of this receptor. Each was used to label the CCK receptor expressed on Chinese hamster ovary-CCKR cells, with the labeled domain then released by cyanogen bromide cleavage. Capillary electrophoresis with laser-induced fluorescence detection achieved an on-capillary mass sensitivity of 1.6 attomoles (10 ؊18 mol), with an excellent signal-to-noise ratio. Each of the biologically divergent, but structurally similar probes saturably and specifically labeled the same receptor domain, consistent with conservation of "docking" determinants. This had an apparent mass of 2.9 kDa, most consistent with the first extracellular loop domain. An additional probe having its site of covalent attachment in a different region of the probe (residue 29 of CCK-33) labeled a distinct receptor fragment with differential migration on capillary electrophoresis (third extracellular loop). Identification of the specific receptor residue(s) covalently linked to the amino-terminal probes must await further fragmentation and sequence analysis.Remarkable diversity exists for structures capable of high affinity binding and activation of G protein-coupled receptors. Understanding the molecular determinants of ligand binding provides important insights that may be useful in structurebased drug design and in understanding the basic mechanisms of receptor activation. Photoaffinity labeling provides the most direct way to identify sites of contact between residues in ligands and a receptor (1); however, traditionally this involves use of large amounts of radioactivity and inefficient and timeconsuming purification procedures. In this work, we developed a simple, rapid, and efficient procedure for analyzing sites of ligand-receptor binding, and applied this to a series of structurally related ligands representing agonists, partial agonist, and antagonist of the CCK receptor.Capillary electrophoresis (CE), 1 initially performed by Jorgenson and Lukacs (2), has emerged in recent years as a versatile and powerful tool for the analysis of biological molecules, because of its unparalleled efficiency for separation, small injection volumes, and low mass detection limits (3, 4). We recently developed a CE method for the separation of peptides cleaved from hydrophobic membrane proteins, such as bacteriorhodopsin, a possible model for the study of G protein-coupled receptors (5). In the present work, we have optimized and extended this to apply to extremely sparse, w...

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“…It can either correspond to the adsorption of particles in the presence of an external field acting parallel to the surface or to the adsorption on an inclined substrate. Two specific examples of the first case are capillary electrophoresis [12] and adsorption in the presence of an imposed shear for sufficiently small diffusion layers [8] [13]. To be precise, we will study the adsorption of disks of diameter one which arrive at the surface forming an angle α with the normal to the line (see Fig.…”

mentioning

confidence: 99%

Adsorption of Colloidal Particles in the Presence of External Fields

1995

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We present a new class of sequential adsorption models in which the adsorbing particles reach the surface following an inclined direction (shadow models). Capillary electrophoresis, adsorption in the presence of a shear or on an inclined substrate are physical manifestations of these models. Numerical simulations are carried out to show how the new adsorption mechanisms are responsible for the formation of more ordered adsorbed layers and have important implications in the kinetics, in particular modifying the jamming limit.

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Capillary Electrophoresis

Cited by 296 publications

References 452 publications

Performance of a physically adsorbed high-molecular-mass polyethyleneimine layer as coating for the separation of basic proteins and peptides by capillary electrophoresis

Performance of a physically adsorbed high-molecular-mass polyethyleneimine layer as coating for the separation of basic proteins and peptides by capillary electrophoresis

Structurally Related Peptide Agonist, Partial Agonist, and Antagonist Occupy a Similar Binding Pocket within the Cholecystokinin Receptor

Adsorption of Colloidal Particles in the Presence of External Fields

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