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
王伊莹 1周小玲 1叶子弘 1张雅芬1
作者信息
- 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
- 折叠
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.
Abstract
Interleukin-11(IL-11)is a multifunctional cytokine that plays a crucial role in various bio-logical processes,including the promotion of hematopoiesis,regulation of the immune system,and facili-tation 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 es-sential.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-dilu-tion concentration and gradient of IL-11,cell inoculum size,and IL-11 incubation duration.Comprehen-sive 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 ap-proach for detecting the biological activity of IL-11.This innovative method significantly enhances the de-tection 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Key words
interleukin-11(IL-11)/reporter gene method/biological activity/detection引用本文复制引用
基金项目
国家重点研发计划(2021YFF0600800)
出版年
2024
NETL
NSTL
万方数据