Nonaqueous capillary zone electrophoresis of synthetic organic polypeptides

Hervé Cottet, Willy Vayaboury, Daniel Kirby, Olivia Giani, Jacques Taillades, François Schué

Research output: Contribution to journalArticle

Abstract

Poly(Nε-trifluoroacetyl-l-lysine) was used as a model solute to investigate the potential of nonaqueous capillary electrophoresis (NACE) for the characterization of synthetic organic polymers. The information obtained by NACE was compared to that derived from size exclusion chromatography (SEC) experiments, and the two techniques were found to be complimentary for polymer characterization. On one hand, NACE permitted (i) the separation of oligomers according to their molar mass and (ii) the separation of the polymers according to the nature of the end groups. On the other hand, SEC experiments were used for the characterization of the molar mass distribution for higher molar masses. Due to the tendency of the solutes (polypeptides) to adsorb onto the fused-silica capillary wall, careful attention was paid to the rinsing procedure of the capillary between runs in order to keep the capillary surface clean. For that purpose, the use of electrophoretic desorption under denaturating conditions was very effective. Optimization of the separation was performed by studying (i) the influence of the proportion of methanol in a methanol/acetonitrile mixture and (ii) the influence of acetic acid concentration in the background electrolyte. Highly resolved separation of the oligomers (up to a degree of polymerization n of ∼50) was obtained by adding trifluoroacetic acid to the electrolyte. Important information concerning the polymer conformations could be obtained from the mobility data. Two different plots relating the effective mobility data to the degree of polymerization were proposed for monitoring the changes in polymer conformations as a function of the number of monomers.
Original languageEnglish
Pages (from-to)5554-5560
Number of pages7
JournalAnalytical Chemistry
Volume75
Issue number20
DOIs
Publication statusPublished - Sep 2003

Fingerprint

Electrophoresis
Capillary electrophoresis
Molar mass
Polymers
Peptides
Size exclusion chromatography
Oligomers
Electrolytes
Methanol
Conformations
Polymerization
Trifluoroacetic Acid
Organic polymers
Fused silica
Acetic Acid
Lysine
Desorption
Monomers
Experiments
Monitoring

Keywords

  • acetates
  • acetic acid
  • acetonitriles
  • gel chromatography
  • capillary electrophoresis
  • light
  • lysine
  • methanol
  • molecular weight
  • peptides
  • polylysine
  • polymers
  • radiation scattering
  • solvents

Cite this

Cottet, H., Vayaboury, W., Kirby, D., Giani, O., Taillades, J., & Schué, F. (2003). Nonaqueous capillary zone electrophoresis of synthetic organic polypeptides. Analytical Chemistry, 75(20), 5554-5560. https://doi.org/10.1021/ac034526o
Cottet, Hervé ; Vayaboury, Willy ; Kirby, Daniel ; Giani, Olivia ; Taillades, Jacques ; Schué, François. / Nonaqueous capillary zone electrophoresis of synthetic organic polypeptides. In: Analytical Chemistry. 2003 ; Vol. 75, No. 20. pp. 5554-5560.
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Cottet, H, Vayaboury, W, Kirby, D, Giani, O, Taillades, J & Schué, F 2003, 'Nonaqueous capillary zone electrophoresis of synthetic organic polypeptides', Analytical Chemistry, vol. 75, no. 20, pp. 5554-5560. https://doi.org/10.1021/ac034526o

Nonaqueous capillary zone electrophoresis of synthetic organic polypeptides. / Cottet, Hervé; Vayaboury, Willy; Kirby, Daniel; Giani, Olivia; Taillades, Jacques; Schué, François.

In: Analytical Chemistry, Vol. 75, No. 20, 09.2003, p. 5554-5560.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Nonaqueous capillary zone electrophoresis of synthetic organic polypeptides

AU - Cottet, Hervé

AU - Vayaboury, Willy

AU - Kirby, Daniel

AU - Giani, Olivia

AU - Taillades, Jacques

AU - Schué, François

PY - 2003/9

Y1 - 2003/9

N2 - Poly(Nε-trifluoroacetyl-l-lysine) was used as a model solute to investigate the potential of nonaqueous capillary electrophoresis (NACE) for the characterization of synthetic organic polymers. The information obtained by NACE was compared to that derived from size exclusion chromatography (SEC) experiments, and the two techniques were found to be complimentary for polymer characterization. On one hand, NACE permitted (i) the separation of oligomers according to their molar mass and (ii) the separation of the polymers according to the nature of the end groups. On the other hand, SEC experiments were used for the characterization of the molar mass distribution for higher molar masses. Due to the tendency of the solutes (polypeptides) to adsorb onto the fused-silica capillary wall, careful attention was paid to the rinsing procedure of the capillary between runs in order to keep the capillary surface clean. For that purpose, the use of electrophoretic desorption under denaturating conditions was very effective. Optimization of the separation was performed by studying (i) the influence of the proportion of methanol in a methanol/acetonitrile mixture and (ii) the influence of acetic acid concentration in the background electrolyte. Highly resolved separation of the oligomers (up to a degree of polymerization n of ∼50) was obtained by adding trifluoroacetic acid to the electrolyte. Important information concerning the polymer conformations could be obtained from the mobility data. Two different plots relating the effective mobility data to the degree of polymerization were proposed for monitoring the changes in polymer conformations as a function of the number of monomers.

AB - Poly(Nε-trifluoroacetyl-l-lysine) was used as a model solute to investigate the potential of nonaqueous capillary electrophoresis (NACE) for the characterization of synthetic organic polymers. The information obtained by NACE was compared to that derived from size exclusion chromatography (SEC) experiments, and the two techniques were found to be complimentary for polymer characterization. On one hand, NACE permitted (i) the separation of oligomers according to their molar mass and (ii) the separation of the polymers according to the nature of the end groups. On the other hand, SEC experiments were used for the characterization of the molar mass distribution for higher molar masses. Due to the tendency of the solutes (polypeptides) to adsorb onto the fused-silica capillary wall, careful attention was paid to the rinsing procedure of the capillary between runs in order to keep the capillary surface clean. For that purpose, the use of electrophoretic desorption under denaturating conditions was very effective. Optimization of the separation was performed by studying (i) the influence of the proportion of methanol in a methanol/acetonitrile mixture and (ii) the influence of acetic acid concentration in the background electrolyte. Highly resolved separation of the oligomers (up to a degree of polymerization n of ∼50) was obtained by adding trifluoroacetic acid to the electrolyte. Important information concerning the polymer conformations could be obtained from the mobility data. Two different plots relating the effective mobility data to the degree of polymerization were proposed for monitoring the changes in polymer conformations as a function of the number of monomers.

KW - acetates

KW - acetic acid

KW - acetonitriles

KW - gel chromatography

KW - capillary electrophoresis

KW - light

KW - lysine

KW - methanol

KW - molecular weight

KW - peptides

KW - polylysine

KW - polymers

KW - radiation scattering

KW - solvents

UR - http://pubs.acs.org/doi/abs/10.1021/ac034526o

U2 - 10.1021/ac034526o

DO - 10.1021/ac034526o

M3 - Article

VL - 75

SP - 5554

EP - 5560

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 20

ER -