Analysis of alpha-1-acid glycoprotein isoforms using CE-LIF with fluorescent thiol derivatization

Raúl Garrido-Medina, Angel Puerta, Zuly Rivera-Monroy, Mercedes de Frutos, Andras Guttman, José Carlos Diez-Masa

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The analysis of glycoprotein isoforms is of high interest in the biomedical field and clinical chemistry. Many studies have demonstrated that some glycoprotein isoforms could serve as biomarkers for several major diseases, such as cancers and vascular diseases, among others. Capillary zone electrophoresis (CZE) is a well-established technique to separate glycoprotein isoforms, however, it suffers from limited sensitivity when UV-Vis detection is used. On the other hand, with laser-induced fluorescence (LIF) detection, derivatization reaction to render the proteins fluorescent can destroy the resolution of the isoforms. In this work, a derivatization procedure through the thiol groups of glycoproteins using either 5-(iodoacetamide) fluorescein (5-IAF) or BODIPY iodoacetamide is presented with the model protein of alpha-1-acid glycoprotein (AGP). The derivatization process presented enabled high-resolution analysis of AGP isoforms by CZE-LIF. The derivatization procedure was successfully applied to label AGP from samples of serum and secretome of artery tissue, enabling the separation of the AGP isoforms by CE-LIF in natural samples at different concentration levels.

Original languageEnglish
Pages (from-to)1113-1119
Number of pages7
JournalELECTROPHORESIS
Volume33
Issue number7
DOIs
Publication statusPublished - Apr 1 2012

Keywords

  • AGP-isoforms
  • BODIPY
  • CZE
  • Fluorescein
  • LIF-detection

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

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  • Cite this

    Garrido-Medina, R., Puerta, A., Rivera-Monroy, Z., de Frutos, M., Guttman, A., & Diez-Masa, J. C. (2012). Analysis of alpha-1-acid glycoprotein isoforms using CE-LIF with fluorescent thiol derivatization. ELECTROPHORESIS, 33(7), 1113-1119. https://doi.org/10.1002/elps.201100473