| Mass spectrometric immunoassay of intact insulin and related variants for population proteomics studies. | |
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MedLine Citation:
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PMID: 21656909 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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PURPOSE: The purpose of the work presented herein was to develop a high-throughput assay for the quantification of human insulin in plasma samples while simultaneously detecting, with high mass accuracy, any additional variant forms of insulin that might be present in each sample. EXPERIMENTAL DESIGN: A mass spectrometric immunoassay (MSIA) was designed in which anti-human insulin antibodies were immobilized to commercially available mass spectrometric immunoassay pipette tips and used to capture insulin and related protein variants from human plasma. RESULTS: Standard curves for insulin exhibited linearity (average R(2) for three days of analysis=0.99) and assay concentration limits of detection and limits of quantification for insulin were found to be 1 and 15 pM, respectively. Estimated coefficient of variations for inter-day experiments (n=3 days) were <8%. Simultaneously, the assay was shown to detect and identify insulin metabolites and synthetic insulin analogs (e.g. Lantus). Notably, insulin variants not known to exist in plasma were detected in diabetics. CONCLUSIONS AND CLINICAL RELEVANCE: This introductory study sets a foundation toward the screening of large populations to investigate insulin isoforms, isoform frequencies, and their quantification. |
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Authors:
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Paul E Oran; Jason W Jarvis; Chad R Borges; Nisha D Sherma; Randall W Nelson |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural Date: 2011-06-08 |
Journal Detail:
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Title: Proteomics. Clinical applications Volume: 5 ISSN: 1862-8354 ISO Abbreviation: Proteomics Clin Appl Publication Date: 2011 Aug |
Date Detail:
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Created Date: 2011-08-01 Completed Date: 2011-11-15 Revised Date: 2012-01-16 |
Medline Journal Info:
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Nlm Unique ID: 101298608 Medline TA: Proteomics Clin Appl Country: Germany |
Other Details:
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Languages: eng Pagination: 454-9 Citation Subset: IM |
Copyright Information:
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Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. |
Affiliation:
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Molecular Biomarkers, The Biodesign Institute at Arizona State University, Tempe, AZ 85287, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Antibodies, Immobilized / immunology, metabolism* Genetics, Population High-Throughput Screening Assays / methods* Humans Immunoassay / methods* Insulin / blood*, genetics, immunology Limit of Detection Mass Spectrometry / methods* Polymorphism, Genetic Protein Isoforms / blood*, genetics, immunology Proteomics / methods* Reference Standards Reproducibility of Results Sensitivity and Specificity Swine |
| Grant Support | |
ID/Acronym/Agency:
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R01 DK082542-03/DK/NIDDK NIH HHS; R01DK082542/DK/NIDDK NIH HHS; R24DK083948/DK/NIDDK NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Antibodies, Immobilized; 0/Insulin; 0/Protein Isoforms |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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