Targeted Selected Reaction Monitoring Mass Spectrometric Immunoassay for Insulin-like Growth Factor 1

被引：38
作者：
Niederkofler, Eric E. [1 ]
Phillips, David A. [1 ]
Krastins, Bryan [2 ]
Kulasingam, Vathany [3 ,4 ]
Kiernan, Urban A. [1 ]
Tubbs, Kemmons A. [1 ]
Peterman, Scott M. [2 ]
Prakash, Amol [2 ]
Diamandis, Eleftherios P. [3 ,4 ]
Lopez, Mary F. [2 ]
Nedelkov, Dobrin [1 ]
机构：
[1] Thermo Fisher Sci, Tempe, AZ USA
[2] Thermo Fisher Sci, BRIMS, Cambridge, MA USA
[3] Univ Hlth Network, Toronto, ON, Canada
[4] Univ Toronto, Toronto, ON, Canada
来源：
PLOS ONE | 2013年 / 8卷 / 11期
关键词：
IGF-I ASSAYS; HORMONE ABUSE; QUANTIFICATION; PLASMA; RADIOIMMUNOASSAY; REPRODUCIBILITY; EXTRACTION; BIOMARKERS; PROTEINS; PEPTIDE;
D O I：
10.1371/journal.pone.0081125
中图分类号：
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号：
07 ; 0710 ; 09 ;
摘要：
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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