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Old 02-09-2020, 06:07 AM
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rss IGF - Development of a novel assay for IGFBP-2 complexed with IGF-I and-II in human s

Development of a novel assay for IGFBP-2 complexed with IGF-I and-II in human serum.

Growth Horm IGF Res. 2020 Jan 21;51:38-45

Authors: Agerholm J, Hjortebjerg R, Espelund U, Rasmussen TR, Folkersen B, Bjerre M, Frystyk J


Abstract
BACKGROUND: Insulin-like growth factor binding-protein 2 (IGFBP-2) was originally identified as an IGF-carrier, governing IGF half-life, tissue accessibility and biological effects. Later, IGFBP-2 was discovered to possess IGF-independent effects. IGFBP-2 circulates in several forms, as free protein, complexed with IGF-I or IGF-II, or as IGFBP-2 fragments. The various IGFBP-2 forms are all included when measuring serum IGFBP-2 concentrations by immunoassay (i.e., immunoreactive (ir-)IGFBP-2). In this study, we describe a novel method to measure the amount of IGF that circulates bound to IGFBP-2.
METHOD: IGFBP-2 was immunoprecipitated from human serum using magnetic beads, which were subsequently eluted by acidification. After neutralization, eluates were assayed for ir-IGFBP-2, IGF-I and IGF-II and compared to serum concentrations. This allowed measurement of IGFBP-2-compexed IGF-I and IGF-II, respectively. To test the method clinically, serum from 146 patients with lung cancer, 151 patients with non-cancer pulmonary diseases and 28 healthy controls were analyzed.
RESULTS: We immuno-precipitated 97.±.3.3% of serum IGFBP-2 and recovered > 75% of IGFBP-2-complexed IGFs, with intra- and inter-assay coefficient of variations (CVs) averaging < 5% and.
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