title: Anti-human fibroblast growth factor-21 monoclonal antibody preparation, characterization and analysis ofin vitrobioactivity.
abstract: Human fibroblast growth factor 21 (hFGF-21) is involved in numerous metabolic processes and elevated hFGF-21 levels are associated with many metabolic diseases. However, the role hFGF-21 serves in the metabolic system is not fully understood. A humanized anti-hFGF-21 monoclonal antibody (mAb) would provide a novel method for further investigations into the role hFGF-21 serves in the metabolic system and related diseases, which may reveal therapeutic targets for future treatment of these diseases. The present study aimed to prepare an anti-hFGF-21 mAb, followed by identification of its characteristics and bioactivityin vitro. The results of the present study identified that the anti-hFGF-21 mAb (clone 2D8) produced had good specificity, had an immunoglobulin isotype of IgG2b and a titer of 1:1.024×106. hFGF-21 was screened for epitopes using fluorescence-activated cell sorting, which revealed a specific 15 amino acid sequence (YQSEAHGLPLHLPGN) that the anti-hFGF-21 mAb recognized.In vitrobioactivity of anti-hFGF-21 was determined using a glucose uptake assay and by measuring the expression of glucose transporter 1 (GLUT1) messenger RNA (mRNA) in 3T3-L1 adipocytes. This revealed that hFGF-21-dependent glucose uptake and GLUT1 mRNA expression were negatively correlated with increasing levels of the anti-hFGF-21 mAb tested, and that hFGF-21 activity could be overcome by increasing concentrations of the mAb, demonstrating that the mAb has hFGF-21-neutralizing activityin vitro.
authors.affiliation: Biopharmaceutical Department, College of Life Science, Northeast Agricultural University, Harbin, Heilongjiang 150030, P.R. China.
authors.forename: Liangjun
authors.initials: L
authors.lastname: Ding
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, Heilongjiang 150069, P.R. China.
authors.forename: Zhichao
authors.initials: Z
authors.lastname: Hao
authors.affiliation: Biopharmaceutical Department, College of Life Science, Northeast Agricultural University, Harbin, Heilongjiang 150030, P.R. China.
authors.forename: Qingyan
authors.initials: Q
authors.lastname: Yuan
authors.affiliation: Biopharmaceutical Department, College of Life Science, Northeast Agricultural University, Harbin, Heilongjiang 150030, P.R. China.
authors.forename: Pengfei
authors.initials: P
authors.lastname: Xu
authors.affiliation: Biopharmaceutical Department, College of Life Science, Northeast Agricultural University, Harbin, Heilongjiang 150030, P.R. China.
authors.forename: Yinhang
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Biopharmaceutical Department, College of Life Science, Northeast Agricultural University, Harbin, Heilongjiang 150030, P.R. China.
authors.forename: Deshan
authors.initials: D
authors.lastname: Li
keywords: bioactivity; characterization; epitope; glucose uptake