Setup for human sera MALDI profiling: The case of rhEPO treatment

Chiara Fania, Michele Vasso, Enrica Torretta, Paul Robach, Gaetano Cairo, Carsten Lundby, Cecilia Gelfi

Research output: Contribution to journalArticle

Abstract

The implementation of high-throughput technologies based on qualitative and quantitative methodologies for the characterization of complex protein mixtures is increasingly required in clinical laboratories. MALDI profiling is a robust and sensitive technology although the serum high dynamic range imposes a major limitation hampering the identification of less abundant species decreasing the quality of MALDI profiling. A setup to improve these parameters has been performed for recombinant human erythropoietin (rhEPO) monitoring in serum, analyzing the effects of two commercially available columns (MARS Hu7 and Hu14) for immunodepletion, and two matrices (α-cyano-4-hydroxycinnamic acid and 2',4'-dihydroxyacetophenone) for peak quality improvement. The immunodepletion capability of both columns was determined by 2-D DIGE, which precisely revealed the efficacy of Hu14 in protein removal and the serum dynamic range decrement. In addition, the type of matrix, the sample dilution, and the efficacy of optimized parameters were used for serum profiling of ten healthy subjects before and after rhEPO treatment. The principal component analysis indicates that a combination of Hu14 column and 2',4'-dihydroxyacetophenone matrix increases data quality allowing the discrimination between treated and untreated samples, making serum MALDI profiling suitable for clinical monitoring of rhEPO.

Original languageEnglish
Pages (from-to)1715-1727
Number of pages13
JournalElectrophoresis
Volume32
Issue number13
DOIs
Publication statusPublished - Jun 2011

    Fingerprint

Keywords

  • 2-D differential gel electrophoresis
  • EPO
  • Immunodepletion
  • MALDI profiling
  • Serum

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

Cite this

Fania, C., Vasso, M., Torretta, E., Robach, P., Cairo, G., Lundby, C., & Gelfi, C. (2011). Setup for human sera MALDI profiling: The case of rhEPO treatment. Electrophoresis, 32(13), 1715-1727. https://doi.org/10.1002/elps.201100134