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抗人白介素-17RA单克隆抗体的制备及活性检测

Preparation and Activity Detection of A Novel Monoclonal Antibody to Human Interleukin-17RA
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摘要 目的制备具有中和人白介素-17RA(h IL-17RA)免疫活性的单克隆抗体,并探讨其生物学活性。方法通过基因工程技术制备抗h IL-17RA的单克隆抗体。采用OctetRED系统检测其亲和力,ELISA、Western blot和流式细胞术分析抗体的结合作用和种属交叉反应,人包皮成纤维细胞-1上细胞因子分泌实验检测抗体的中和活性。结果抗h IL-17RA单克隆抗体亲和力常数KD<1.0×10^(-12)mol/L,并与其他物种有一定的交叉结合反应,它能结合白介素(IL)-17RA并有效阻断其与配体的结合,抑制下游相关细胞因子IL-6、IL-8和人CXC趋化因子配体1的分泌。结论成功制备1株抗h IL-17RA的中和抗体,为IL-17RA相关疾病通路的研究提供了工具,也为IL-17RA靶点药物的开发奠定了一定的基础。 Objective To develop a neutralizing antibody to human interleukin-17RA( h IL-17RA)and investigate the biological activity of the antibody. Methods The anti-h IL-17 RA antibody was produced by recombinant technology while its binding affinity to h IL-17 RA was measured by OctetRED system. The antibody's blocking activity to h IL-17 RA and cross-reaction between species were analyzed by enzyme-linked immunosorbent assay,Western blot,fluorescence activated cell sorter,while its neutralization activity was measured by inhibiting the secretion of cytokine in human foreskin fibroblast cells-1. Results The antibody's affinity constant was less than 1. 0 × 10^(- 12) mol/L, with certain cross-reaction among species. Secretion of interleukin( IL)-6,IL-8,and chemokine( CXC motif) ligand 1 elicited by IL-17 A,IL-17 A/F,and IL-17 F in human foreskin fibroblast cells-1 was neutralized by the antibody. Conclusion A novel neutralizing antibody to h IL-17 RA was obtained,which provides research tools for studying h IL-17 RA signal pathways and lays a foundation for drug development targeting h IL-17 RA.
出处 《中国医学科学院学报》 CAS CSCD 北大核心 2016年第4期428-433,共6页 Acta Academiae Medicinae Sinicae
基金 十二五"重大新药创制"科技重大专项(2013ZX09402301)~~
关键词 白介素-17RA 白介素-17A 白介素-17F 抗体 人包皮成纤维细胞-1 interleukin-17RA interleukin-17A interleukin-17F antibody human foreskin fibroblast cells-1
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