Use of fullerene-, octadecyl-, and triaconthyl silica for solid phase extraction of tryptic peptides obtained from unmodified and in vitro glycated human serum albumin and fibrinogen

Katalin Böddi, Anikó Takátsy, Szilvia Szabó, Lajos Markó, L. Márk, I. Wittmann, R. Ohmacht, Gergely Montskó, Rainer M. Vallant, Thomas Ringer, Rania Bakry, Christian W. Huck, Günther K. Bonn, Zoltán Szabó

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

SPE plays a crucial role in bioanalytical research. In the present work a novel fullerene(C60)-derivatised silica material is compared with octadecyl(C18) - and triaconthyl(C30)-silicas regarding recoveries of peptides and sequence coverage of HSA and fibrinogen digests. C30- and C60(30 nm)-SPE materials were found to be the two most prominent SPE materials. At low peptide concentrations C60-material prepared from a silica gel with a pore size of 30 nm has proven to be the best material with regards to recoveries. By increasing the amount of loaded peptides recoveries decrease due to its relative low binding capacity in contrast to C30-silica particles, showing no changes. The best sequence coverages of Aα-and Bβ-chains of 20 pmol fibrinogen digest can also be achieved using these two SPE materials, C60 (30 nm) demonstrates an outstanding value of sequence coverage (62.15%) achieved for the γ-chain. After nonenzymatic glycation the digests of fibrinogen and HSA were also separated. This makes the detection of a considerably higher number of glycated peptides possible compared to the unfractionated digests and the use of boronate affinity chromatography in the case of fibrinogen. For HSA, ten new sites of glycation at lysine and arginine residues have been explored. Using the detailed SPE/offline MALDI method the glycation sites on fibrinogen are first described in this paper.

Original languageEnglish
Pages (from-to)295-308
Number of pages14
JournalJournal of Separation Science
Volume32
Issue number2
DOIs
Publication statusPublished - Jan 2009

Fingerprint

Fullerenes
Serum Albumin
Silicon Dioxide
Fibrinogen
Peptides
Silica
Recovery
Affinity chromatography
Arginine
Silica Gel
Silica gel
Lysine
Pore size

Keywords

  • Fullerene
  • Glycation
  • MALDI-TOF
  • Pepties
  • RP-materials

ASJC Scopus subject areas

  • Analytical Chemistry
  • Filtration and Separation

Cite this

Use of fullerene-, octadecyl-, and triaconthyl silica for solid phase extraction of tryptic peptides obtained from unmodified and in vitro glycated human serum albumin and fibrinogen. / Böddi, Katalin; Takátsy, Anikó; Szabó, Szilvia; Markó, Lajos; Márk, L.; Wittmann, I.; Ohmacht, R.; Montskó, Gergely; Vallant, Rainer M.; Ringer, Thomas; Bakry, Rania; Huck, Christian W.; Bonn, Günther K.; Szabó, Zoltán.

In: Journal of Separation Science, Vol. 32, No. 2, 01.2009, p. 295-308.

Research output: Contribution to journalArticle

Böddi, Katalin ; Takátsy, Anikó ; Szabó, Szilvia ; Markó, Lajos ; Márk, L. ; Wittmann, I. ; Ohmacht, R. ; Montskó, Gergely ; Vallant, Rainer M. ; Ringer, Thomas ; Bakry, Rania ; Huck, Christian W. ; Bonn, Günther K. ; Szabó, Zoltán. / Use of fullerene-, octadecyl-, and triaconthyl silica for solid phase extraction of tryptic peptides obtained from unmodified and in vitro glycated human serum albumin and fibrinogen. In: Journal of Separation Science. 2009 ; Vol. 32, No. 2. pp. 295-308.
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AU - Markó, Lajos

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AU - Montskó, Gergely

AU - Vallant, Rainer M.

AU - Ringer, Thomas

AU - Bakry, Rania

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