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Efficient and reproducible analysis of peptides by capillary electrophoresis using noncovalently bilayer-coated capillaries.
Electrophoresis. 2004 Mar; 25(6):817-24.E

Abstract

The usefulness of a noncovalent capillary coating consisting of two layers of oppositely charged polymers for the separation of peptides with capillary electrophoresis (CE) was studied. Capillaries were coated simply by subsequently flushing with solutions of 1% m/v Polybrene and 1% v/v poly(vinylsulfonate) (PVS) forming a bilayer, which showed to produce a strong and highly reproducible electroosmotic flow (EOF) at low pH. Using this coating in combination with a background electrolyte (BGE) containing sodium phosphate (pH 2.5) and 0.01% v/v PVS, initially broadened and overlapping peaks were obtained for some test peptides. By omitting the PVS from the BGE, the peak width and shape of the peptides improved resulting in baseline separation. A systematic study of the influence of the BGE composition showed that considerable further enhancement of the separation efficiency was achieved by increasing the ionic strength of the BGE. Using a BGE of 200 mM tris(hydroxymethyl)aminomethane (Tris)-phosphate (pH 2.5) plate numbers for the peptides were in the 300 000-600 000 range and the relative standard deviation of the peptide migration times was less then 0.3% (n = 5). The use of Tris-phosphate instead of sodium phosphate allowed the current to stay within acceptable limits when 30 kV was used as separation voltage. Overall, the bilayer coating showed a remarkable EOF repeatability, as well as long-term stability. Compared to bare fused-silica capillaries the intraday and interday repeatability of migration times was very favorable and coated capillaries could be used for over a month performing analyses with low and high ionic strength BGEs without any performance deterioration. The usefulness of the bilayer-coated capillaries for the analysis of positively charged peptides was demonstrated by the fast and efficient separation of various closely related enkephalins and the baseline separation of an isomeric peptide/peptoid couple exhibiting efficiencies of over 550 000 plates.

Authors+Show Affiliations

Catai JR
Department of Biomedical Analysis, Utrecht University, NL-3508 TB Utrecht, The Netherlands.
Somsen GW
No affiliation info available
de Jong GJ
No affiliation info available

MeSH

ElectrolytesElectrophoresis, CapillaryHexadimethrine BromidePeptidesPolyvinylsReproducibility of ResultsSulfonic Acids

Pub Type(s)

Journal Article

Language

eng

PubMed ID

15004841

Citation

Catai, Jonatan R., et al. "Efficient and Reproducible Analysis of Peptides By Capillary Electrophoresis Using Noncovalently Bilayer-coated Capillaries." Electrophoresis, vol. 25, no. 6, 2004, pp. 817-24.
Catai JR, Somsen GW, de Jong GJ. Efficient and reproducible analysis of peptides by capillary electrophoresis using noncovalently bilayer-coated capillaries. Electrophoresis. 2004;25(6):817-24.
Catai, J. R., Somsen, G. W., & de Jong, G. J. (2004). Efficient and reproducible analysis of peptides by capillary electrophoresis using noncovalently bilayer-coated capillaries. Electrophoresis, 25(6), 817-24.
Catai JR, Somsen GW, de Jong GJ. Efficient and Reproducible Analysis of Peptides By Capillary Electrophoresis Using Noncovalently Bilayer-coated Capillaries. Electrophoresis. 2004;25(6):817-24. PubMed PMID: 15004841.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Efficient and reproducible analysis of peptides by capillary electrophoresis using noncovalently bilayer-coated capillaries. AU - Catai,Jonatan R, AU - Somsen,Govert W, AU - de Jong,Gerhardus J, PY - 2004/3/9/pubmed PY - 2004/11/2/medline PY - 2004/3/9/entrez SP - 817 EP - 24 JF - Electrophoresis JO - Electrophoresis VL - 25 IS - 6 N2 - The usefulness of a noncovalent capillary coating consisting of two layers of oppositely charged polymers for the separation of peptides with capillary electrophoresis (CE) was studied. Capillaries were coated simply by subsequently flushing with solutions of 1% m/v Polybrene and 1% v/v poly(vinylsulfonate) (PVS) forming a bilayer, which showed to produce a strong and highly reproducible electroosmotic flow (EOF) at low pH. Using this coating in combination with a background electrolyte (BGE) containing sodium phosphate (pH 2.5) and 0.01% v/v PVS, initially broadened and overlapping peaks were obtained for some test peptides. By omitting the PVS from the BGE, the peak width and shape of the peptides improved resulting in baseline separation. A systematic study of the influence of the BGE composition showed that considerable further enhancement of the separation efficiency was achieved by increasing the ionic strength of the BGE. Using a BGE of 200 mM tris(hydroxymethyl)aminomethane (Tris)-phosphate (pH 2.5) plate numbers for the peptides were in the 300 000-600 000 range and the relative standard deviation of the peptide migration times was less then 0.3% (n = 5). The use of Tris-phosphate instead of sodium phosphate allowed the current to stay within acceptable limits when 30 kV was used as separation voltage. Overall, the bilayer coating showed a remarkable EOF repeatability, as well as long-term stability. Compared to bare fused-silica capillaries the intraday and interday repeatability of migration times was very favorable and coated capillaries could be used for over a month performing analyses with low and high ionic strength BGEs without any performance deterioration. The usefulness of the bilayer-coated capillaries for the analysis of positively charged peptides was demonstrated by the fast and efficient separation of various closely related enkephalins and the baseline separation of an isomeric peptide/peptoid couple exhibiting efficiencies of over 550 000 plates. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/15004841/Efficient_and_reproducible_analysis_of_peptides_by_capillary_electrophoresis_using_noncovalently_bilayer_coated_capillaries_ DB - PRIME DP - Unbound Medicine ER -