基于报告基因法检测白细胞介素-11生物学活性  

Detection of the Biological Activity of Interleukin-11 Based on the Reporter Gene Method

作　　者：王伊莹 周小玲 叶子弘 张雅芬 WANG Yi-Ying;ZHOU Xiao-Ling;YE Zi-Hong;ZHANG Ya-Fen(Zhejiang Provincial Key Laboratory of Biometrology and Inspection&Quarantine,College of Life Sciences,China Jiliang University,Hangzhou 310018,China)

机构地区：[1]中国计量大学生命科学学院,浙江省生物计量及检验检疫技术重点实验室,杭州310018

出　　处：《中国生物化学与分子生物学报》2024年第10期1462-1470,共9页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家重点研发计划(No.2021YFF0600800)资助。

摘　　要：白细胞介素-11(interleukin-11,IL-11)是一种多功能细胞因子,具有促进造血、免疫调节和组织修复等多种功能,在医学研究和临床治疗中具有重要的应用价值,因此,IL-11生物学活性的准确检测和评估至关重要。现采用的细胞增殖抑制法操作繁琐、耗时较长和易受到白细胞介素-6影响。本研究首先根据IL-11的作用机制(JAK-STAT信号通路),构建可稳定响应IL-11的萤光素酶报告基因表达单克隆细胞株,然后对IL-11的预稀释浓度及稀释梯度、接种细胞量和IL-11孵育时间进行优化,进一步对方法的准确性、精密度、特异性、稳定性以及和药典方法一致性进行全面验证,建立了基于报告基因法检测IL-11生物学活性的新方法。该方法可有效提高检测准确性和灵敏度,并且缩短检测时间,具有一定的应用前景。Interleukin-11(IL-11)is a multifunctional cytokine that plays a crucial role in various biological processes,including the promotion of hematopoiesis,regulation of the immune system,and facilitation of tissue repair.These functions highlight its significant importance in both medical research and clinical therapy.Consequently,the precise detection and assessment of IL-11′s biological activity are essential.The currently employed cell proliferation inhibition assay is cumbersome,time-consuming,and susceptible to interference from Interleukin-6.Here we study the JAK-STAT signaling pathway,a key mechanism of IL-11 actions.We initially establish a stable monoclonal cell line that expresses a luciferase reporter gene responsive to IL-11.We subsequently optimized critical parameters,including the pre-dilution concentration and gradient of IL-11,cell inoculum size,and IL-11 incubation duration.Comprehensive validation was undertaken to evaluate accuracy,precision,specificity,stability,and concordance with pharmacopeial methods,ultimately leading to the establishment of a novel reporter gene-based approach for detecting the biological activity of IL-11.This innovative method significantly enhances the detection accuracy and sensitivity while reducing the testing time,thereby offering promising prospects for broad applications.

关 键 词：白介素-11 报告基因法 生物学活性 检测 

分 类 号：Q511[生物学—生物化学]