Abstract |
Insulin-like growth factor 1 (IGF1) is an important biomarker of human growth disorders that is routinely analyzed in clinical laboratories. Mass spectrometry-based workflows offer a viable alternative to standard IGF1 immunoassays, which utilize various pre-analytical preparation strategies. In this work we developed an assay that incorporates a novel sample preparation method for dissociating IGF1 from its binding proteins. The workflow also includes an immunoaffinity step using antibody-derivatized pipette tips, followed by elution, trypsin digestion, and LC-MS/MS separation and detection of the signature peptides in a selected reaction monitoring (SRM) mode. The resulting quantitative mass spectrometric immunoassay (MSIA) exhibited good linearity in the range of 1 to 1,500 ng/mL IGF1, intra- and inter-assay precision with CVs of less than 10%, and lowest limits of detection of 1 ng/mL. The linearity and recovery characteristics of the assay were also established, and the new method compared to a commercially available immunoassay using a large cohort of human serum samples. The IGF1 SRM MSIA is well suited for use in clinical laboratories.
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Authors | Eric E Niederkofler, David A Phillips, Bryan Krastins, Vathany Kulasingam, Urban A Kiernan, Kemmons A Tubbs, Scott M Peterman, Amol Prakash, Eleftherios P Diamandis, Mary F Lopez, Dobrin Nedelkov |
Journal | PloS one
(PLoS One)
Vol. 8
Issue 11
Pg. e81125
( 2013)
ISSN: 1932-6203 [Electronic] United States |
PMID | 24278387
(Publication Type: Journal Article)
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Chemical References |
- Biomarkers
- Insulin-Like Growth Factor I
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Topics |
- Biomarkers
(blood)
- Humans
- Immunoassay
(methods, standards)
- Insulin-Like Growth Factor I
(chemistry, metabolism)
- Mass Spectrometry
(methods, standards)
- Reference Standards
- Reference Values
- Reproducibility of Results
- Sensitivity and Specificity
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