在Leishmania tarentolae中应用2A-系统用于多基斯特龙人体IgG的产生
Jing Yi Lai1, Kae Yi Tan2, Choo Hock Tan3
1Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 Penang, Malaysia.
Methods (San Diego, Calif.)
|December 17, 2025
概括
莱什马尼亚tarentolae提供了一个具有成本效益的平台表达单克隆抗体,包括人类scFv-Fc和IgG格式. 这种真核细胞系统允许N-链糖化,使其成为昂贵的哺乳动物细胞系的可行替代品.
科学领域:
- 生物技术和生物加工
- 蛋白质表达系统是一种蛋白质表达系统.
- 免疫学 免疫学 免疫学
背景情况:
- 哺乳动物细胞系是单克隆抗体表达的标准,原因是翻译后的修改.
- 哺乳动物表达系统是昂贵的,需要专门的基础设施.
- 在Leishmania tarentolae中,真核体表达提供了一个具有成本效益的替代方案,即N-链接的糖化.
研究的目的:
- 评估Leishmania tarentolae作为单克隆抗体的表达平台.
- 为了表达人类IgG抗体,对L. tarentolae.中Naja kaouthia长链神经毒素 (LNTX) 进行表达.
- 评估表达抗体的产量,同质性和完整性.
主要方法:
- 从菌体显示库中对LNTX进行人类单克隆抗体scFv的分离.
- 将scFv转换为scFv-Fc和全长的IgG格式.
- 在L. tarentolae中使用基于2A的二基斯特龙载体表达IgG.
- 分析IgG表达产量,同质性和降解.
主要成果:
- 在L. tarentolae.中成功表达人类IgG对LNTX.
- 通过二基斯特龙向量系统实现了良好的表达收益率.
- 表达的IgG在尺寸和最小降解方面呈现同质性.
- L. tarentolae 显示出表达人类 scFv-Fc 和 IgG 抗体的潜力.
结论:
- 莱什马尼亚tarentolae是表达人类单克隆抗体的可行和经济有效的替代方案.
- 该平台支持N相关的糖化,这对抗体功能至关重要.
- 这项研究验证了L. tarentolae能够产生复杂的抗体格式,如IgG.
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