阴优化调节IgG3和IgM的表达和糖化在N. 班塔米亚纳 (Benthamiana) 是一种植物
Lin Sun1, Somanath Kallolimath1, Roman Palt1
1Department of Applied Genetics and Cell Biology, Institute of Plant Biotechnology and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Austria.
Frontiers in bioengineering and biotechnology
|December 21, 2023
概括
在植物中优化DNA编码子的使用,可以增强人类单克隆抗体 (mAbs) 的产生. 然而,这种增加的表达可以改变糖化,可能会影响治疗疗效和免疫性.
科学领域:
- 植物生物技术 植物生物技术
- 分子生物学分子生物学
- 免疫学 免疫学 免疫学
背景情况:
- 植物正在成为生产复杂人体蛋白质的平台,如单克隆抗体 (mAbs).
- 优化DNA编码子是增强植物重组蛋白表达的关键策略.
研究的目的:
- 为了研究DNA编码子优化对尼科蒂亚娜本塔米亚纳人类单克隆抗体 (mAbs) 暂时表达的影响.
- 为了评估生物化学性质和子优化 (CO) 与非子优化 (NCO) mAbs. 的糖化模式.
主要方法:
- 在Nicotiana benthamiana中,针对SARS-CoV-2的三个人类mAbs (一个IgG3,两个IgM) 的过渡表达.
- 对码头优化 (CO) 和非码头优化 (NCO) 变体的比较.
- 分析使用qRT-PCR检测mRNA水平,SDS-PAGE检测蛋白质纯度,以及抗原结合试验.
主要成果:
- 在所有测试的mAbs.中,Codon优化显著增加了mRNA水平和总体蛋白质表达.
- 纯化CO和NCOmAbs在基本生化特性或抗原结合方面没有显著差异.
- 增强的表达导致改变了糖化,特别是增加了N-糖甘加工中的曼诺基基结构.
结论:
- 优化DNA编码子是有效的促进植物中的重组 mAb 生产.
- 增加的表达水平可以微妙地改变糖化样式,影响糖占用率和N-糖甘加工.
- 仔细监测重组蛋白质至关重要,特别是在治疗应用中,因为糖化变化可能会影响疗效和免疫性.
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