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Mass Spectrometric Immunoassay for Insulin‐Like Growth Factor 1
Author(s) -
Nedelkov Dobrin,
Niederkofler Eric,
Phillips David,
Krastins Bryan,
Kiernan Urban,
Tubbs Kemmons,
Lopez Mary
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.997.2
Subject(s) - chromatography , analyte , immunoassay , chemistry , elution , insulin like growth factor , normalization (sociology) , pipette , antibody , growth factor , biochemistry , biology , receptor , sociology , anthropology , immunology
Mass Spectrometric Immunoassays (MSIA) are protein quantification methods that utilize mass spectrometric detection for quantification of targeted protein analytes affinity retrieved by surface‐immobilized antibodies. This rather straightforward concept is realized through affinity pipette devices (MSIA‐Tips) that enable high‐throughput assaying of hundreds of samples per day. Described here is the development and validation of mass spectrometric immunoassay for insulin‐like growth factor 1 (IGF1). MSIA‐Tips derivatized with anti‐IGF‐1 antibody were used for affinity isolation of IGF1 from human samples. Long R3 (LR3) IGF1, spiked into samples prior to the affinity isolation, served as internal reference standard for signal normalization and IGF1 quantification. Following capture and elution, both the intact proteins and their surrogate tryptic peptides were analyzed via LC/MS. Linear standard curves were built that spanned the range of 1–1,500 ng/mL. The assay exhibited intra‐ and inter‐assay precisions of <10%, and linearity and spiking‐recovery in the 90–110% range. Data was compared to that obtained with a commercially available ELISA, yielding good correlation. The data correlation between intact IGF1 and peptide‐based IGF1 MS analyses was good, demonstrating the applicability of the IGF1 MSIA across MS platforms.