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Derivatized graphitic nanofibres (GNF) as a new support material for mass spectrometric analysis of peptides and proteins

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Abstract

Graphitic nanofibres (GNFs), 100–200 nm in diameter and 5–20 μm in length have been modified in order to yield different affinities (Cu2+ and Fe3+ loaded immobilized metal affinity chromatography (IMAC) as well as cation and anion exchange materials) for the extraction of a range of biomolecules by their inherited hydrophobicity and the hydrophilic chemical functionalities, obtained by derivatization. Modified GNFs have for the first time been employed as carrier materials for protein profiling in material-enhanced laser desorption/ionization (MELDI) for the enrichment and screening of biofluids. For that purpose, the derivatized GNF materials have comprehensively been characterized regarding surface area, structural changes during derivatization, IMAC, as well as ion exchange and protein-loading capacity and recovery. GNF derivatives revealed high protein-binding capacity (2,000 μg ml−1 for insulin) and ideal sensitivities, resulting in a detection limit of 50 fmol μl−1 (for insulin), which is crucial for the detection of low abundant species in biological samples. Compared to other MELDI carrier materials, sensitivity was enhanced on GNF derivatives, which might be ascribed to the fact that GNFs support desorption and ionization mechanisms and by absorbing laser energy in addition to matrix.

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Acknowledgments

This work was supported by the Genome Research in Austria (GEN-AU) (Federal Ministry for Education, Science and Culture, Vienna, Austria), by the West Austrian Initiative for Nano Networking (WINN) and by the Austrian Science Foundation (FWF), SFB Project 021 (Vienna, Austria).

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  1. Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University, Innrain 52a, 6020, Innsbruck, Austria

    Andreas Greiderer, Matthias Rainer, Muhammad Najam-ul-Haq, Rainer M. Vallant, Christian W. Huck & Günther K. Bonn

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  1. Andreas Greiderer
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  2. Matthias Rainer
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  3. Muhammad Najam-ul-Haq
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  4. Rainer M. Vallant
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  5. Christian W. Huck
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  6. Günther K. Bonn
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Correspondence to Christian W. Huck.

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Greiderer, A., Rainer, M., Najam-ul-Haq, M. et al. Derivatized graphitic nanofibres (GNF) as a new support material for mass spectrometric analysis of peptides and proteins. Amino Acids 37, 341–348 (2009). https://doi.org/10.1007/s00726-008-0159-x

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  • DOI: https://doi.org/10.1007/s00726-008-0159-x

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