Cation-exchange chromatography of peptides on poly(2-sulfoethyl aspartamide)-silica

Alpert, Andrew J.; Andrews, Philip C.

doi:10.1016/s0021-9673(00)94785-x

Cited by 132 publications

(71 citation statements)

References 27 publications

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“…For this purpose, cation-exchange or anion-exchange chromatography using a salt gradient have previously been investigated for separating basic and acidic peptides, respectively (Dizdaroglu, 1985;Isobe et al, 1982;Mant and Hodges, 1985;Takahashi et al, 1983). A more general IEC method has been reported by Alpert and Andrews (1988), who used cation-exchange chromatography with a mobile phase pH of 3.0. The method takes advantage of the fact that at pH 3 peptides lose their negative charges at aspartic and glutamic acid residues and the C-termini, and therefore become positively charged due to the presence of basic amino acid residues and the N-termini, so that nearly all peptides will adsorb onto a cation-exchange column packing.…”

Section: General Considerationsmentioning

confidence: 99%

“…Although these two methods were moderately successful in separating peptides, neither of the methods exploits the characteristics of true chromatofocusing, where a pH gradient is formed dynamically inside the column so that focusing effects are maximized. The IEC method developed by Alpert and Andrews (1988) discussed above would similarly appear to offer opportunities for improvement in some situations since the separation achieved is based only on the difference in the charges at the basic residues. It is therefore reasonable to expect that chromatofocusing may have advantages in certain applications over previously investigated ion-exchange methods for separating peptides.…”

Section: General Considerationsmentioning

confidence: 99%

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Chromatofocusing of peptides and proteins using linear pH gradients formed on strong ion‐exchange adsorbents

Kang

Frey

2004

Biotech & Bioengineering

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Although it is commonly believed that a column packing used for chromatofocusing must have an ''even'' buffering capacity in order to produce a linear pH gradient, it is demonstrated here that linear pH gradients suitable for chromatofocusing can be produced on a column packing having a minimal buffering capacity. In particular, if either a strong-acid cation-exchange column packing or a strongbase anion-exchange column packing is presaturated with either a weak acid titrated with a strong base, or a weak base titrated with a strong acid, respectively, to the initial pH, then a linear or nearly linear pH gradient can be formed using a polyampholyte elution buffer by taking advantage of the presence of small quantities of weak-acid or weakbase functional groups that generally exist on these types of column packings. Experimental and theoretical studies are used to demonstrate that such systems have potential advantages over traditional chromatofocusing methods in terms of the speed of the separation, the resolution achieved, and the range of applications possible. Among other techniques described, a method for separating tryptic peptides using chromatofocusing and a strong-acid cation-exchange column packing is demonstrated to be a useful alternative to capillary isoelectric focusing and ionexchange chromatography using a salt gradient for this purpose. B

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Section: General Considerationsmentioning

confidence: 99%

Section: General Considerationsmentioning

confidence: 99%

Chromatofocusing of peptides and proteins using linear pH gradients formed on strong ion‐exchange adsorbents

Kang

Frey

2004

Biotech & Bioengineering

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“…The separations show complementary selectivity to RP systems (Alpert and Andrews, 1988). The separation efficiency may be poor in mobile phases containing less than 90% acetonitrile; hence the PolySulfoethyl A column is recommended for not too strongly retained samples (Hartmann et al, 2003).…”

Section: Polysuccinimide Bonded Stationary Phasesmentioning

confidence: 99%

Theory and Practice of UHPLC and UHPLC–MS

Guillarme

Veuthey

2017

Handbook of Advanced Chromatography /Mass Spectrometry Techniques

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“…Similarly, polymeric reverse phase materials (PLRP) offers increased mechanical strength, uniform hydrophobicity, and high recovery and PLRP materials have also been utilized widely for the intact protein separations [27,29,30]. In contrast to RPLC, hydrophobic interaction liquid chromatography (HILIC) uses a polar stationary phase and gradients increasing water content which results in the elution of more hydrophobic species first [31,32]. Modified histone forms separation using HILIC prior to top-down MS has been reported [33,34].…”

Section: Reverse Phase Liquid Chromatography [Rplc]mentioning

confidence: 99%

Top-down proteomics: applications, recent developments and perspectives

Pamreddy

Panyala

2016

J Appl Bioanal

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REVIEWNow-a-days, top-down proteomics (TDP) is a booming approach for the analysis of intact proteins and it is attaining significant interest in the field of protein biology. The term has emerged as an alternative to the well-established, bottom-up strategies for analysis of peptide fragments derived from either enzymatically or chemically digestion of intact proteins. TDP is applied to mass spectrometric analysis of intact large biomolecules that are constituents of protein complexes and assemblies. This article delivers an overview of the methodologies in top-down mass spectrometry, mass spectrometry instrumentation and an extensive review of applications covering the venomics, biomedical research, protein biology including the analysis of protein post-translational modifications (PTMs), protein biophysics, and protein complexes. In addition, limitations of top-down proteomics, challenges and future directions of TDP are also discussed.

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Cation-exchange chromatography of peptides on poly(2-sulfoethyl aspartamide)-silica

Cited by 132 publications

References 27 publications

Chromatofocusing of peptides and proteins using linear pH gradients formed on strong ion‐exchange adsorbents

Chromatofocusing of peptides and proteins using linear pH gradients formed on strong ion‐exchange adsorbents

Theory and Practice of UHPLC and UHPLC–MS

Top-down proteomics: applications, recent developments and perspectives

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