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Updated: Aug 16, 2026

Murine Model of CD40-activation of B cells

Murine Model of CD40-activation of B cells

Published on: March 5, 2010

在 prokaryotic 系统中,复合溶性 CD40 配体的表达和功能性特征

Qi Xu¹, Xinying He¹, Jiale Bai¹

¹Key Laboratory of Brain Functional Genomics (East China Normal University), Ministry of Education, Shanghai Key Laboratory of Brain Functional Genomics (East China Normal University), School of Life Science, East China Normal University.

Protein expression and purification

|December 20, 2025

概括

一个优化的 prokaryotic 系统有效地产生功能性可溶性 CD40 连接体 (sCD40L). 这种重组sCD40L增强了血小板聚合,减少了出血时间,提供了治疗潜力.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
血液学血液学血液学

背景情况:

可溶性CD40连接体 (sCD40L) 在免疫反应和血液静止中起作用.
功能性sCD40L的高效生产对于研究和治疗开发至关重要.

研究的目的:

建立一个优化的 prokaryotic 表达系统,以产生功能性可溶性 CD40 连接体 (sCD40L).
描述重组sCD40L的静血活性.

主要方法:

编码优化的sCD40L基因合成并克隆到pET28a载体中.
在由IPTG诱导的大肠杆菌BL21(DE3) 中的表达.
通过亲和染色法进行净化;通过SDS-PAGE和西方 blot进行分析.
通过体外血小板聚合和体内血液静止试验评估的生物活性.

主要成果:

成功构建了pET28a-sCD40L表达向量.
达到>90%的蛋白质纯度,通过SDS-PAGE和西部斑点 (∼20kDa) 得到证实.
重组sCD40L在小鼠中显示了剂量依赖的血小板聚合增强和显著减少出血时间.

结论:

优化的 prokaryotic 系统有效地产生功能性的 sCD40L 无需eukaryotic 转化后修改.
为研究CD40L介导的血液静止提供了一个有价值的工具.
开发针对血静的治疗应用的潜力.

关键词:

埃舍里希亚大肠杆菌 (Escherichia coli) 是一个大肠杆菌.血液静止血液静止血液静止血小板聚合是血小板的聚合.Prokaryotic 的表达方式可溶性CD40连接物 (sCD40L) 的存在.

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