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.
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Authors | Matthias Rainer, Muhammad Najam-ul-Haq, Rania Bakry, Christian W Huck, Günther K Bonn |
Journal | Journal of proteome research
(J Proteome Res)
Vol. 6
Issue 1
Pg. 382-6
(Jan 2007)
ISSN: 1535-3893 [Print] United States |
PMID | 17203982
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Biomarkers
- Blood Proteins
- Chelating Agents
- Imino Acids
- Peptides
- Polymethacrylic Acids
- Polyvinyls
- poly(glycidyl methacrylate-co-divinylbenzene)
- Copper
- iminodiacetic acid
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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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