| Quantification of protein calibrants by amino acid analysis using isotope dilution mass spectrometry. | |
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MedLine Citation:
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PMID: 20807498 Owner: NLM Status: In-Process |
Abstract/OtherAbstract:
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This work demonstrates that amino acid analysis based on isotope dilution mass spectrometry (IDMS) can be applied to quantify proteins having different complexities and natures. Five proteins and one decapeptide were selected for the study: C-reactive protein (CRP), beta-2-microglobulin (B2M), cystatine C (CysC), human serum albumin (HSA), Ara h1, and angiotensin I. The quantification was based on the determination of four amino acids, proline (Pro), isoleucine (Ile), valine (Val), and phenylalanine (Phe) within a working range between 5 and 100 pmol/injection of each amino acid, after 60 min digestion with HCl at 150°C. The amino acids were selected taking into account their abundance in the protein sequence and to include the more difficult to break peptide bonds. Quantification of the protein amounts calculated from each amino acid is consistent, indicating that the method is working reliably. This consistency points to a complete hydrolysis of the proteins. The trueness of the method was proven when dry mass determination after dialysis was applied to HSA and CRP and the results were compared to those from amino acid analysis. Traceability to SI was assured by extensive characterisation of the amino acid calibrants by nuclear magnetic resonance, neutron activation analysis, and Karl Fischer titration. |
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Authors:
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A Muñoz; R Kral; H Schimmel |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-08-31 |
Journal Detail:
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Title: Analytical biochemistry Volume: 408 ISSN: 1096-0309 ISO Abbreviation: Anal. Biochem. Publication Date: 2011 Jan |
Date Detail:
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Created Date: 2010-10-26 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0370535 Medline TA: Anal Biochem Country: United States |
Other Details:
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Languages: eng Pagination: 124-31 Citation Subset: IM |
Copyright Information:
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Copyright © 2010 Elsevier Inc. All rights reserved. |
Affiliation:
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Joint Research Centre, Institute for Reference Material and Measurements, Geel, Belgium. amalia.MUNOZ-PINEIRO@ec.europa.eu |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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