Influence of pH on formation and stability of phosphatidylcholine/phosphatidylserine coatings in fused-silica capillaries

Hautala, Jari T.; Wiedmer, Susanne Κ.; Riekkola, Marja‐Liisa

doi:10.1002/elps.200406143

Cited by 29 publications

(28 citation statements)

References 18 publications

(12 reference statements)

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“…Since then, we have chosen HEPES (at pH 7.4) as BGE solution in most of our liposome studies. The importance of the piperazine ring in HEPES was recognized and its role as a linking agent between the fused-silica wall and the anionic phospholipid membrane was studied in detail [6]. Protonation of the amino groups in HEPES and the polar head groups of the phospholipids enhanced the attachment of phospholipids to the silica wall.…”

Section: Introductionmentioning

confidence: 99%

Piperazine-based buffers for liposome coating of capillaries for electrophoresis

et al. 2005

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Anionic liposomes can be coated on fused-silica capillaries for electrophoresis in the presence of N-(hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) (HEPES) as background electrolyte (BGE) solution. In this work, the interaction of various compounds with zwitterionic and anionic phospholipid coatings was studied with HEPES at pH 7.4 as BGE solution. The chromatographic and electrophoretic behavior of three test sample solutions (anionic, cationic, and neutral) was investigated for evaluation of the phospholipid coatings. Our results show that hydrophobic interactions between analytes and the phospholipid coating are important for the migration of charged analytes. In addition, the performances of other piperazine-based buffers, i.e., N-(2-hydroxyethyl)piperazine-N'-(2-hydroxypropanesulfonic acid), piperazine-N,N'-bis(2-ethanesulfonic acid), and piperazine-N,N'-bis(hydroxypropane sulfonic acid), at pH 7.4, as liposome solvent and BGE solution were evaluated and compared with the performance of HEPES at pH 7.4. The anionic liposome solution comprised 80/20 mol% phosphatidylcholine/phosphatidylserine. A simple test solution was selected and the chromatographic and electrophoretic migration behavior of the analytes was evaluated. The results show that, in addition to HEPES, other piperazine-based buffers at pH 7.4 are suitable for coating of fused-silica capillaries with anionic liposomes.

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Section: Introductionmentioning

confidence: 99%

Piperazine-based buffers for liposome coating of capillaries for electrophoresis

et al. 2005

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“…1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (another analogue of DLPC with a longer hydrophobic chain), and a mixture of 1-palmitoyl-2-oleoylsn-glycero-3-phosphocholine and the anionic phosphatidylserine were used to form capillary coatings for the separations of standard basic proteins and steroids [20,21].…”

Section: Introductionmentioning

confidence: 99%

Sphingomyelins as semi‐permanent capillary coatings for protein separations in CE and off‐line analysis with MALDI‐MS

Jurčić

Yeung

2009

Electrophoresis

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Phosphatidylcholine (PC) is one of the major phospholipids that make up the biological cell membrane. It was previously reported to form capillary inner wall coatings for CE. The zwitterionic head group of PC produced a neutral net-charged bilayer, which was found effective in preventing wall adsorption of both cationic and anionic proteins [J. M. Cunliffe et al. Anal. Chem. 2002, 74, 776-783]. Another major membrane phospholipid that possesses a zwitterionic head group is sphingomyelin (SM). In this work, the novel characterization of SM on its effectiveness in capillary coating formation for CE separations of proteins and peptides was presented. Similar properties were observed between PC and SM, including their effects on the EOF, peak efficiencies, and migration time reproducibilities. SM appeared to be more readily soluble in aqueous solutions, and it was found equally effective as PC in facilitating protein separation. The main difference observed was their performances in delivering a peptide mixture for off-line analysis with MALDI-MS. Superior sample recovery was evident from the capillary coated with SM compared with that with PC. The number of peptides identified from a 1 ng/microL myglobin tryptic digest sample increased from 5 to 16 (42 and 69%, respectively in protein sequence coverage).

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“…In addition, the strong avidin-biotin binding applied in this study should enhance the stability of the avidin immobilization in the membrane. In previous studies we found that addition of PS to phosphatidylcholine (PC)-containing phospholipids vesicles enhances separations in liposome EKC (LEKC) owing to the increased negative charge [28] and in CEC owing to the greater number of amines taking part in the immobilization of membrane on the silica wall [29].…”

Section: Resultsmentioning

confidence: 99%

“…We assume that, owing to the high molar mass of avidin, there are a large number of hydrophobic amino acids present to stabilize the phospholipid coatings still further. In previous work we recognized the importance of the piperazine ring in Hepes, not only as a linking agent between the fusedsilica wall and the anionic phospholipid membrane but also as stabilizer of the coating [29]. Protonation of the amino groups in HEPES and the polar head groups of the phospholipids enhanced the attachment of phospholipids to the silica wall.…”

Section: Effect Of Buffer Liposome Composition and Extra Avidin Stementioning

confidence: 97%

Immobilization of phospholipid‐avidin on fused‐silica capillaries for chiral separation in open‐tubular capillary electrochromatography

et al. 2006

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Phospholipid-coated fused-silica capillaries with immobilized avidin were applied in the chiral separation of D,L-tryptophan, D,L-PTH-serine, and D,L-PTH-threonine at pH 7.4 by open-tubular CEC. Liposomes prepared from 1,2-dipalmitoyl-sn-glycero-3-phosphocholine, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(Cap biotinyl), or 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(Biotinyl) with different amounts of phosphatidylserine were assessed as phospholipid coating materials. The stability of the coating and the success of the coating procedure were evaluated in terms of the repeatability of the enantiomer migration times and the resolution of enantiomers. The coating procedure itself significantly affected the migration times and resolution of the enantiomers. Reliable chiral separations with high separation efficiencies were achieved through careful choice of the coating method.

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Influence of pH on formation and stability of phosphatidylcholine/phosphatidylserine coatings in fused-silica capillaries

Cited by 29 publications

References 18 publications

Piperazine-based buffers for liposome coating of capillaries for electrophoresis

Piperazine-based buffers for liposome coating of capillaries for electrophoresis

Sphingomyelins as semi‐permanent capillary coatings for protein separations in CE and off‐line analysis with MALDI‐MS

Immobilization of phospholipid‐avidin on fused‐silica capillaries for chiral separation in open‐tubular capillary electrochromatography

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