Capillary electrophoresis of synthetic peptide standards varying in charge and hydrophobicity

Popa, Traian; Mant, Colin T.; Hodges, Robert S.

doi:10.1002/elps.200305576

Cited by 13 publications

(5 citation statements)

References 33 publications

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“…FS capillary covalently coated with C8-RP or by physical adsorption coated with hydroxyethyl cellulose has been used for separation of synthetic peptide standards varying in charge and hydrophobicity [171]. Several (metallo)-porphyrins, particularly the porphyrin derivative tetraphenylporphyrin, and complexes of porphyrin derivatives with metal ions (Zn [173].…”

Section: Electrochromatographymentioning

confidence: 99%

Recent developments in capillary electrophoresis and capillary electrochromatography of peptides

2006

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The article gives a comprehensive review on the recent developments in the applications of high-performance capillary electromigration methods, zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography, to analysis, preparation, and physicochemical characterization of peptides. The article presents new approaches to the theoretical description and experimental verification of electromigration behavior of peptides, covers the methodological aspects of capillary electroseparations of peptides, such as rational selection of separation conditions, sample preparation, suppression of peptide adsorption, new developments in individual separation modes, and new designs of detection systems. Several types of applications of capillary electromigration methods to peptide analysis are presented: conventional qualitative and quantitative analysis, purity control, determination in biomatrices, monitoring of chemical and enzymatical reactions and physical changes, amino acid and sequence analysis, and peptide mapping of proteins. Some examples of micropreparative peptide separations are given and capabilities of capillary electromigration techniques to provide important physicochemical characteristics of peptides are demonstrated.

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Section: Electrochromatographymentioning

confidence: 99%

Recent developments in capillary electrophoresis and capillary electrochromatography of peptides

2006

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“…4 and 5), migration times of the peptides increase with increasing hydrophobicity of the acidic counterion (TFA -< PFPA -< HFBA -).The separations at these high levels of perfluorinated acid concentrations represent bidimensional separations: (i) peptides of the same length but of different nominal charge are separated in order of decreasing charge (i.e., +3 peptides migrate the earliest and +1 peptides migrate the last) according to a CZE mechanism; (ii) within each group of peptides, the peptides are separated in order of increasing hydrophobicity according to an hydrophobically-mediated mechanism introduced by the anionic ion-pairing reagent. The method which achieves this bidimensional separation is referred to as Ion-Interaction-Capillary Zone Electrophoresis (II-CZE) [30,37,38]. Clearly, for this mixture of 27 peptide standards, RP-HPLC (based solely/ mainly on overall peptide hydrophobicity) in the presence of 20 mM H 3 PO 4 or 20 mM TFA (typical RP-HPLC conditions for peptide separations) (Fig.…”

Section: Ce Of Peptidesmentioning

confidence: 99%

Ion-interaction–capillary zone electrophoresis of cationic proteomic peptide standards

Popa¹,

Mant

Hodges

2006

Journal of Chromatography A

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We have employed a novel capillary electrophoresis (CE) approach recently developed in our laboratory, termed ion-interaction-capillary zone electrophoresis (II-CZE), to the resolution of a mixture of 27 synthetic cationic proteomic peptide standards. These peptides were comprised of three groups of nine peptides (with net charges of +1, +2 and +3 for all nine peptides within a group), the hydrophobicity of the nine peptides within a group varying only subtly between adjacent peptides. This bidimensional CE approach achieved excellent resolution of the peptides with high peak capacity by combining the powerful CZE mechanism located in the background electrolyte (BGE) with an hydrophobicity-based mechanism also located in the BGE, the latter consisting of high concentrations (up to 0.4 M) of aqueous perfluorinated acids ( trifluoroacetic acid, pentafluoropropionic acid and heptafluorobutyric acid). Thus, concomitant with a CZE separation of the three differently charged groups of peptides, there is an hydrophobically-mediated separation of the peptides within these groups effected through interaction of the hydrophobic anions of the perfluorinated acids with hydrophobic amino acid side-chains in the peptides. This methodology is dramatically different from other CE methods that have used complexing agents such as micelles or cyclodextrins in MEKC. Overall, the results presented here demonstrate the value of CE as a peptide separative tool in its own right, including its use for proteomic applications, and not merely as a complementary technique to reversed-phase high-performance liquid chromatography (RP-HPLC).

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“…In addition, the buffer pH was lowered to pH 2.0 in order to increase the positive charge of the peptides and subsequently ion-ion interactions between the positively charged peptides and the negatively charged perfluorinated acids. This approach has also been termed "ion-interaction CZE" [18]. As a general trend, the separation of the peptide diastereomers improved with increasing hydrophobicity (chain length) and concentration of the perfluorinated acids.…”

Section: Cementioning

confidence: 98%

Recent advances in peptide and peptidomimetic stereoisomer separations by capillary electromigration techniques

Scriba¹

2006

Electrophoresis

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As the stereochemistry of peptides determines their physicochemical properties and biological activities, analytical methods able to discriminate between peptide stereoisomers are important especially with regard to pharmaceutical peptides and peptidomimetics. The present review summarizes recent developments in peptide and peptidomimetic stereoisomer separations by capillary electromigration techniques. The majority of separations were performed by CE while only few reports have been published on the subject of electrochromatography. In addition to systematic studies on the applicability of certain buffer additives and the evaluation of specific experimental conditions, there have been attempts to understand the mechanistic aspects of peptide stereoisomer separations as well as to analyze the structure of peptide-CD complexes.

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Capillary electrophoresis of synthetic peptide standards varying in charge and hydrophobicity

Cited by 13 publications

References 33 publications

Recent developments in capillary electrophoresis and capillary electrochromatography of peptides

Recent developments in capillary electrophoresis and capillary electrochromatography of peptides

Ion-interaction–capillary zone electrophoresis of cationic proteomic peptide standards

Recent advances in peptide and peptidomimetic stereoisomer separations by capillary electromigration techniques

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