Ce hydrogen deuterium exchange-ms in peptide analysis

S.S.M. Lau, N.M. Stainforth, D.G. Watson, G.G. Skellern, S.A.C. Wren, J.N.A. Tettey

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

CE and hydrogen-deuterium (H/D) exchange MS are useful tools in the analysis and characterisation of peptides. This study reports the facile coupling of these tools in the H/D exchange CE-MS analysis of model and pharmaceutically important peptides, using a sheath flow interface. The peptides varied in mass from 556 (leucine enkephalin) to 1620 Da (bombesin), and in charge state from 0.33 (leucine enkephalin) to 3.0 (substance P). The application of a BGE composed of ammonium formate buffer (25 mM, pD 3.5 in D2O (>98% D atom)), a sheath liquid composed of formic acid (0.25% v/v in D2O) and ACN (30:70 v/v), and dissolving the samples in a mixture of ACN/D2O (50:50 v/v) facilitates complete H/D exchange. Because of complete H/D exchange the ESI mass spectra produced are easy to interpret and comparable to those obtained from LC-MS analysis. The CE-H/D-MS approach has the advantage of requiring lower volumes of deuterated solvents. The b- and y-series fragments produced by using in-source decomposition correspond to those predicted. With the peptides studied, the complete exchange H/D exchange observed with both the molecular and fragment ions helps to confirm both amino acid composition and sequence.
LanguageEnglish
Pages393-400
Number of pages7
JournalElectrophoresis
Volume29
Issue number2
DOIs
Publication statusPublished - Jan 2008

Fingerprint

Deuterium
Hydrogen
formic acid
Peptides
Leucine Enkephalin
Bombesin
Substance P
Amino Acid Sequence
Buffers
Ions
Decomposition
Amino Acids
Atoms
Liquids
Chemical analysis

Keywords

  • CE
  • hydrogen-deuterium exchange
  • MS

Cite this

Lau, S. S. M., Stainforth, N. M., Watson, D. G., Skellern, G. G., Wren, S. A. C., & Tettey, J. N. A. (2008). Ce hydrogen deuterium exchange-ms in peptide analysis. Electrophoresis, 29(2), 393-400. https://doi.org/10.1002/elps.200700368
Lau, S.S.M. ; Stainforth, N.M. ; Watson, D.G. ; Skellern, G.G. ; Wren, S.A.C. ; Tettey, J.N.A. / Ce hydrogen deuterium exchange-ms in peptide analysis. In: Electrophoresis. 2008 ; Vol. 29, No. 2. pp. 393-400.
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Lau, SSM, Stainforth, NM, Watson, DG, Skellern, GG, Wren, SAC & Tettey, JNA 2008, 'Ce hydrogen deuterium exchange-ms in peptide analysis' Electrophoresis, vol. 29, no. 2, pp. 393-400. https://doi.org/10.1002/elps.200700368

Ce hydrogen deuterium exchange-ms in peptide analysis. / Lau, S.S.M.; Stainforth, N.M.; Watson, D.G.; Skellern, G.G.; Wren, S.A.C.; Tettey, J.N.A.

In: Electrophoresis, Vol. 29, No. 2, 01.2008, p. 393-400.

Research output: Contribution to journalArticle

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AU - Stainforth, N.M.

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Lau SSM, Stainforth NM, Watson DG, Skellern GG, Wren SAC, Tettey JNA. Ce hydrogen deuterium exchange-ms in peptide analysis. Electrophoresis. 2008 Jan;29(2):393-400. https://doi.org/10.1002/elps.200700368