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

文献类型： 外文期刊

第一作者： Ding, Liangjun

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

作者机构：

关键词： characterization;epitope;glucose uptake;bioactivity

期刊名称：EXPERIMENTAL AND THERAPEUTIC MEDICINE （ 影响因子：2.447； 五年影响因子：2.263 ）

ISSN： 1792-0981

年卷期： 2016 年 12 卷 5 期

页码：

收录情况： SCI

摘要： 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: 1.024x10(6). 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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