Towards stable diagnostic setups in clinical proteomics: Absolute quantitation of peptide biomarkers using MALDI‐TOF‐MS |
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Authors: | Daniel Baechle Katrin Sparbier Hassan Dihazi Sabine Blaschke Gerhard‐Anton Mueller Markus Kostrzewa Thomas Flad Dr |
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Affiliation: | 1. PANATecs GmbH, Tübingen, Germany;2. Bruker Daltonik GmbH, Leipzig, Germany;3. Department of Nephrology and Rheumatology, University Hospital G?ttingen, G?ttingen, Germany;4. PANATecs GmbH, Tübingen, GermanyPANATecs GmbH, Vor dem Kreuzberg 17, 72070 Tübingen, Germany Fax: +49‐7071‐920‐5829 |
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Abstract: | In routine clinical diagnostics, peptide biomarkers are most commonly quantified using immunological techniques but these methods often lack sensitivity and/or specificity. Hence, quantitative mass spectrometry detection is desirable as an alternative diagnostic tool. To date, quantitative mass spectrometry is mostly based on ESI‐MS coupled to LC, requiring highly sophisticated instrumentation and knowledge and is time consuming and expensive. In contrast, MALDI‐TOF‐MS is a very simple, sensitive and rapid method for the detection of peptide biomarkers. However, the infeasibility of absolute quantification has been a tremendous handicap to the use of MS in stable clinical diagnostics. Here, we describe the development of a technical platform based on ClinProt particles and heavy‐isotope internal peptide standards for the fast and reliable preparation of samples. This combines the advantages of MALDI‐TOF as a read‐out system with absolute quantitation of peptide biomarkers. As a proof‐of‐concept, this platform was successfully employed for the absolute determination of the concentration of the highly abundant serum peptide des‐Ala‐Fibrinopeptide A in 45 serum samples from healthy donors. Such technology essentially contributes to the development of a stable MALDI‐TOF‐MS‐based clinical assay. |
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Keywords: | Absolute quantitation Diagnostics MALDI‐TOF‐MS Peptide biomarker |
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