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Targeted selected reaction monitoring mass spectrometric immunoassay for insulin-like growth factor 1.

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.
AuthorsEric 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
JournalPloS one (PLoS One) Vol. 8 Issue 11 Pg. e81125 ( 2013) ISSN: 1932-6203 [Electronic] United States
PMID24278387 (Publication Type: Journal Article)
Chemical References
  • Biomarkers
  • Insulin-Like Growth Factor I
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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