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PLoS One. 2013 Nov 21;8(11):e81125. doi: 10.1371/journal.pone.0081125. eCollection 2013.

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

Author information

1
Thermo Fisher Scientific, Tempe, Arizona, United States of America.

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.

PMID:
24278387
PMCID:
PMC3836743
DOI:
10.1371/journal.pone.0081125
[Indexed for MEDLINE]
Free PMC Article

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