Hybrid phospholipid bilayer coatings for separations of cationic proteins in capillary zone electrophoresis

Gallagher, Elyssia S.; Adem, Seid M.; Bright, Leonard K.; Calderon, Isen Andrew C.; Mansfield, Elisabeth; Aspinwall, Craig A.

doi:10.1002/elps.201300537

Cited by 11 publications

(9 citation statements)

References 36 publications

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“…When compared to other phospholipid‐coated CZE separations of myoglobin, the efficiencies are comparable to those of Gallagher et al. but surpassed by the results from Wang and Lucy . The one notable change in the electrophoresis of myoglobin arises with the cobalt buffer.…”

Section: Resultsmentioning

confidence: 52%

Metal cation control of electroosmotic flow magnitude in phospholipid‐coated capillaries

2016

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CZE has become widespread for the separation and analysis of biomolecules such as proteins and peptides, due to factors such as, the speed of the separations, low sample volume, and high resolution associated with the technique. However, the separation of biomolecules by CZE does present a significant challenge due to the electrostatic attraction and adsorption of cationic, or cation containing, biomolecules to the capillary surface. To that end numerous methods have been developed to passivate, or protect the surface, in order to prevent the adsorption of analytes. Yet, in the process of protecting the capillary surface, the potential for further modification of the EOF, a factor crucial to effective analyte resolution, is greatly diminished. In seeking to overcome this limitation we have explored the potential of incorporating a range of metal cations into a phospholipid bilayer capillary coating. It has previously been established that the inclusion of calcium into the separation buffer with a phospholipid coating will reverse the EOF in the capillary. Here, we present our investigation of a broader range of metal cations included in the separation buffer (Ca(2+) , Mg(2+) , Co(2+) , Ni(2+) , Sr(2+) , Ba(2+) , and Ce(3+) ) revealing that the choice of metal cation can drastically influence the EOF, with observed values between -3.80 × 10(-4) and -5.74 × 10(-5) cm(2) /V·s.

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Section: Resultsmentioning

confidence: 52%

Metal cation control of electroosmotic flow magnitude in phospholipid‐coated capillaries

2016

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“…Supported lipid bilayers have been prepared on a range of substrates including planar silica substrates and curved micro and nanoparticle substrates. Previously, SUVs of bis-SorbPC were fused with planar silica substrates [15, 16], curved capillary surfaces [29, 30, 39], and ca. 100 nm silica nanoparticles [40] to form supported, polymerized phospholipid bilayers, suggesting that fusing bis-SorbPC SUVs with silica microparticles will yield a supported bilayer structure.…”

Section: Resultsmentioning

confidence: 99%

“…Previous studies with FM1-43 have illustrated high levels of fluorescence after intercalation of the dye into lipid membranes [29, 39, 41]. For comparison, both bis-SorbPC and DOPC, a natural lipid, were used to evaluate the stability of bilayer coatings.…”

Section: Resultsmentioning

confidence: 99%

Highly stabilized, polymer–lipid membranes prepared on silica microparticles as stationary phases for capillary chromatography

Gallagher

Adem

Baker

et al. 2015

Journal of Chromatography A

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The ability to rapidly screen complex libraries of pharmacological modulators is paramount to modern drug discovery efforts. This task is particularly challenging for agents that interact with lipid bilayers or membrane proteins due to the limited chemical, physical, and temporal stability of conventional lipid-based chromatographic stationary phases. Here, we describe the preparation and characterization of a novel stationary phase material composed of highly stable, polymeric-phospholipid bilayers self-assembled onto silica microparticles. Polymer lipid membranes were prepared by photochemical or redox initiated polymerization of 1,2-bis[10-(2′,4′-hexadieoyloxy)decanoyl]-sn-glycero-2-phosphocholine (bis-SorbPC), a synthetic, polymerizable lipid. The resulting polymerized bis-SorbPC (poly(bis-SorbPC)) stationary phases exhibited enhanced stability compared to particles coated with 1,2-dioleoyl-sn-glycero-phosphocholine (unpolymerized) phospholipid bilayers when exposed to chemical (50mM triton X-100 or 50% acetonitrile) and physical (15 min sonication) insults after 30 days of storage. Further, poly(bis-SorbPC)-coated particles survived slurry packing into fused silica capillaries, compared to unpolymerized lipid membranes, where the lipid bilayer was destroyed during packing. Frontal chromatographic analyses of the lipophilic small molecules acetylsalicylic acid, benzoic acid, and salicylic acid showed > 44% increase in retention times (P < 0.0001) for all analytes on poly(bis-SorbPC)-functionalized stationary phase compared to bare silica microspheres, suggesting a lipophilic retention mechanism. Phospholipid membrane-functionalized stationary phases that withstand the chemical and physical rigors of capillary LC conditions can substantially increase the efficacy of lipid membrane affinity chromatography, and represents a key advance towards the development of robust membrane protein-functionalized chromatographic stationary phases.

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“…24 In order to enhance the coating stability, the utility of hybrid phospholipid bilayers (HPBs) was studied and applied for the separation of cationic proteins. 51 HPBs are formed by covalently modifying a support with a hydrophobic monolayer into which a self-assembled lipid monolayer is deposited. The authors employed 3-cyanopropyldimethylchlorosilane (CPDCS) or n -octyldimethylchlorosilane (ODCS) for modification of the capillary surface to provide a nominally hydrophobic surface upon which lipid fusion could occur.…”

Section: Separation Modes and Conditionsmentioning

confidence: 99%

Recent advances in protein analysis by capillary and microchip electrophoresis

Dawod

Arvin

Kennedy

2017

Analyst

120

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This review article describes the significant recent advances in analysis of proteins by capillary and microchip electrophoresis during the period mid 2014 to early 2017. The review highlights the progressions, new methodologies, innovative instrumental modifications, and challenges for efficient protein analysis in human specimens, animal tissues, and plant samples. Protein analysis fields covered in this review include analysis of native, reduced, and denatured proteins in addition to Western blotting, protein therapeutics and proteomics.

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Hybrid phospholipid bilayer coatings for separations of cationic proteins in capillary zone electrophoresis

Cited by 11 publications

References 36 publications

Metal cation control of electroosmotic flow magnitude in phospholipid‐coated capillaries

Metal cation control of electroosmotic flow magnitude in phospholipid‐coated capillaries

Highly stabilized, polymer–lipid membranes prepared on silica microparticles as stationary phases for capillary chromatography

Recent advances in protein analysis by capillary and microchip electrophoresis

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