Separation of Peptides and Proteins by Capillary Electrochromatography

Mistry, Kavita; Grinberg, Nelu

doi:10.1081/jlc-120030601

Cited by 12 publications

(5 citation statements)

References 86 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…CEC, which is often described as a hybrid of CZE and LC [6,7], has demonstrated higher separation efficiencies and flow permeabilities than capillary HPLC [8][9][10][11][12][13][14][15][16][17][18][19][20][21][22]. The use of silica-based stationary phases with reversed-phase properties in CEC is well established at medium and higher mobile phase pH, where dissociated residual silanol groups provide the necessary surface charge for a suitable EOF.…”

Section: Introductionmentioning

confidence: 99%

Electrochromatographic retention of peptides on strong cation‐exchange stationary phases

Nischang¹,

Höltzel²,

Tallarek³

2010

Electrophoresis

View full text Add to dashboard Cite

We analyze the systematic and substantial electrical field-dependence of electrochromatographic retention for four counterionic peptides ([Met5]enkephalin, oxytocin, [Arg8]vasopressin, and luteinizing hormone releasing hormone (LHRH) ) on a strong cation-exchange (SCX) stationary phase. Our experiments show that retention behavior in the studied system depends on the charge-selectivity of the stationary phase particles, the applied voltage, and the peptides' net charge. Retention factors of twice positively charged peptides ([Arg8]vasopressin and LHRH at pH 2.7) decrease with increasing applied voltage, whereas lower charged peptides (oxytocin and [Met5]enkephalin at pH 2.7, [Arg8]vasopressin and LHRH at pH 7.0) show a concomitant increase in their retention factors. The observed behavior is explained on the basis of electrical fieldinduced concentration polarization (CP) that develops around the SCX particles of the packing. The intraparticle concentration of charged species (buffer ions, peptides) increases with increasing applied voltage due to diffusive backflux from the enriched CP zone associated with each SCX particle. For twice charged and on the SCX phase strongly retained peptides the local increase in mobile phase ionic strength reduces the electrostatic interactions with the stationary phase, which explains the decrease of retention factors with increasing applied voltage and CP intensity. Lower charged and weaker retained peptides experience a much stronger relative intraparticle enrichment than the twice-charged peptides, which results in a net increase of retention factors with increasing applied voltage. The CP-related contribution to electrochromatographic retention of peptides on the SCX stationary phase is modulated by the applied voltage, the mobile phase ionic strength, and the peptides' net charge and could be used for selectivity tuning in difficult separations.

show abstract

Section: Introductionmentioning

confidence: 99%

Electrochromatographic retention of peptides on strong cation‐exchange stationary phases

Nischang¹,

Höltzel²,

Tallarek³

2010

Electrophoresis

View full text Add to dashboard Cite

show abstract

“…Reversed phase chromatography seems to be more robust and the sample matrix, in contrast to CEC, less influences retention and resolution. Mistry and Grinberg [177] emphasized the Fig. 8.…”

Section: Capillary Electrochromatographymentioning

confidence: 97%

Chromatographic and electrophoretic characterization of protein variants

Ahrer

Jungbauer

2006

Journal of Chromatography B

View full text Add to dashboard Cite

“…Proteins possess various charges and hydrophobic characteristics at different pH values, and thus they can be effectively resolved by CEC using packed beds, monolithics, and open-tubular (OT) stationary phases [160][161][162]. Differences in their charge to volume ratios allow proteins to be separated electrokinetically.…”

Section: Partitionmentioning

confidence: 99%

Capillary electrophoresis‐based separation techniques for the analysis of proteins

Huang

et al. 2006

Electrophoresis

View full text Add to dashboard Cite

CE offers the advantages of high speed, great efficiency, as well as the requirement of minimum amounts of sample and buffer for the analysis of proteins. In this review, we summarize the CE-based techniques coupled with absorption, LIF, and MS detection systems for the analysis of proteins mostly within the past 5 years. The basic principle of each technique and its advantages and disadvantages for protein analysis are discussed in brief. Advanced CE techniques, including on-column concentration techniques and high-efficiency multidimensional separation techniques, for high-throughput protein profiling of complex biological samples and/or of single cells are emphasized. Although the developed techniques provide improved peak capacity, they have not become practical tools for proteomics, mainly because of poor reproducibility, low-sample lading capacity, and low throughput due to ineffective interfaces between two separation dimensions and that between separation and MS systems. In order to identify the complexities and dynamics of the proteomes expressed by cells, tissues, or organisms, techniques providing improved analytical sensitivity, throughput, and dynamic ranges are still demanded.

show abstract

Separation of Peptides and Proteins by Capillary Electrochromatography

Cited by 12 publications

References 86 publications

Electrochromatographic retention of peptides on strong cation‐exchange stationary phases

Electrochromatographic retention of peptides on strong cation‐exchange stationary phases

Chromatographic and electrophoretic characterization of protein variants

Capillary electrophoresis‐based separation techniques for the analysis of proteins

Contact Info

Product

Resources

About