Anti-human fibroblast growth factor-21 monoclonal antibody preparation, characterization and analysis of in vitro bioactivity

Ding,Liangjun; Hao,Zhichao; Yuan,Qingyan; Xu,Pengfei; Yu,Yinhang; Li,Deshan

doi:10.3892/etm.2016.3753

Anti-human fibroblast growth factor-21 monoclonal antibody preparation, characterization and analysis of in vitro bioactivity

Authors:
- Liangjun Ding
- Zhichao Hao
- Qingyan Yuan
- Pengfei Xu
- Yinhang Yu
- Deshan Li
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Affiliations: Biopharmaceutical Department, College of Life Science, Northeast Agricultural University, Harbin, Heilongjiang 150030, P.R. China, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, Heilongjiang 150069, P.R. China
Published online on: September 27, 2016 https://doi.org/10.3892/etm.2016.3753
Pages: 3417-3424

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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 bioactivity in 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 1:1.024x106. 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 vitro bioactivity 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 activity in vitro.

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