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在CHO和HEK细胞系中表达的SARS-CoV-2尖端蛋白质的糖谱比较

Helen L Wright¹, Caroline Evans¹, Philip J Jackson²

¹School of Chemical, Materials and Biological Engineering, University of Sheffield, Mappin St., Sheffield, S1 3JD, UK.

Molecular biotechnology

|October 1, 2024

概括

了解HEK和CHO细胞中产生的SARS-CoV-2尖端蛋白的糖化差异对于开发诊断和疫苗至关重要. CHO细胞产生更复杂的甘氨酸,包括潜在的免疫原体,影响抗原设计.

科学领域:

* 生物化学和分子生物学
* 免疫学和疫苗开发
* 蛋白质工程和生产

背景情况:

*SARS-CoV-2尖端蛋白对于病毒进入至关重要,也是诊断和治疗的主要目标.
* 尖端蛋白的糖化对其功能和免疫性有显著影响.
* 糖化模式的变化源于不同的重组蛋白表达系统,影响产品质量和疗效.

研究的目的:

* 为了比较全长三元型SARS-CoV-2尖端蛋白在人类胚胎 (HEK Expi293F) 和中国仓鼠卵巢 (CHO-S) 细胞中表达的N-糖化概况.
* 用质谱和甘氨酸数据库来描述甘氨基化差异.
* 评估这些糖基化变异对抗原开发和质量控制的影响.

主要方法:

* 在HEK Expi293F和CHO-S细胞系中表达全长三元型SARS-CoV-2尖端蛋白.
*使用双重质谱工作流程进行糖酸分析.
*利用BioPharma与HEK和CHO甘氨酸数据库进行全面的蛋白质表征.

主要成果:

* 在HEK和CHO表达尖端蛋白之间的22个糖化位点中观察到N-甘氨酸类型和多样性的显著差异.
*CHO-S衍生的尖端蛋白显示终端化增加,以及N-glycolylneuraminic acid在一个关键部位的存在.
*与人类细胞 (39个甘氨酸) 相比,CHO细胞具有更大,更复杂的甘氨酸谱 (182个甘氨酸).

结论:

*表达系统的选择深刻影响SARS-CoV-2尖端蛋白质的糖化,影响其功能性和免疫性质.
*CHO衍生的尖端蛋白的独特的糖化概况,特别是化和免疫性甘氨酸,需要对治疗应用进行仔细考虑.
*研究结果支持设计质量 (QbD) 和多属性监测 (MAM) 进行精确的蛋白型分析,有助于开发下一代诊断和针对新出现的SARS-CoV-2变种的疫苗.

关键词:

在N-glycosylation的过程中.在CHO细胞中,在 COVID-19 疫情中,这就是HEK细胞.这就是SARS-CoV-2病毒.

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Conjugated Proteins

Conjugated Proteins

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