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Mass spectrometric identification of serum peptides employing derivatized poly(glycidyl methacrylate/divinyl benzene) particles and mu-HPLC.

Abstract
Biomarkers play a key role in preclinical screening and diagnosis of a disease. Various support materials are utilized for this task, in combination with MALDI-TOF-MS. The way to effectively bind serum contents and their profiling is well-elaborated by the material-enhanced laser desorption ionization (MELDI) approach. In this particular work, focus is placed on the development of a strategy to identify low molecular weight serum peptides. Poly(GMA/DVB) is derivatized in a way to achieve an affinity termed as immobilized metal ion affinity chromatography (IMAC). Iminodiacetic acid (IDA) is used as a chelating ligand, whereas copper (Cu2+) acts as a metal ion for complexing peptides and proteins out of blood serum. Polymer binds the serum compounds over a broad mass range, which includes low mass peptides and high mass albumin (66 kDa). Bound contents are eluted from material by an acetonitrile/trifluoroacetic acid mixture, which proves the reversible nature of metal and amino acid linkage. Polystyrene/divinyl benzene (PS/DVB) monolithic capillary column is used for fractionation through RP-HPLC, prior to the target spotting. The tandem TOF fragment ion mass spectra of each fraction is acquired and used to search against the Swiss-Prot database, using the Mascot search engine for the identification of peptides.
AuthorsMatthias Rainer, Muhammad Najam-ul-Haq, Rania Bakry, Christian W Huck, Günther K Bonn
JournalJournal of proteome research (J Proteome Res) Vol. 6 Issue 1 Pg. 382-6 (Jan 2007) ISSN: 1535-3893 [Print] United States
PMID17203982 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Biomarkers
  • Blood Proteins
  • Chelating Agents
  • Imino Acids
  • Peptides
  • Polymethacrylic Acids
  • Polyvinyls
  • poly(glycidyl methacrylate-co-divinylbenzene)
  • Copper
  • iminodiacetic acid
Topics
  • Amino Acid Sequence
  • Biomarkers (chemistry)
  • Blood Proteins (chemistry)
  • Chelating Agents (pharmacology)
  • Chromatography, High Pressure Liquid (methods)
  • Copper (chemistry)
  • Humans
  • Imino Acids (chemistry)
  • Mass Spectrometry (methods)
  • Molecular Sequence Data
  • Peptides (blood, chemistry)
  • Polymethacrylic Acids (chemistry)
  • Polyvinyls (chemistry)
  • Proteomics (methods)
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

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