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Ricky Lieu¹, Grace Chao¹, Emma Kennedy¹

¹Biotechnology Discovery Research, Eli Lilly and Company, Lilly Biotechnology Center, San Diego, California, USA.

Biotechnology progress

|December 14, 2023

概括

制造稳定,功能性的细胞外域 (ECD) 抗原用于治疗性抗体开发是具有挑战性的. 一种使用抗体碎片作为陪伴者的新型联合表达方法成功生成了难以表达的ECD抗原.

科学领域:

生物化学生物化学
分子生物学分子生物学
蛋白质工程是指蛋白质工程.

背景情况:

细胞外域 (ECD) 抗原对于治疗性抗体的开发至关重要,包括发现,体外检测和表位图绘制.
使用常规表达系统产生功能性和稳定的ECD抗原往往是具有挑战性的,阻碍了治疗进步.

研究的目的:

开发和展示一种有效的方法来产生高质量的,生物活性细胞外域 (ECD) 抗原,特别是那些难以表达的抗原.
为了克服复杂抗原标准蛋白质生产技术的局限性.

主要方法:

一种新的共同表达策略,涉及过渡性共同表达抗原ECD与稳定单克隆抗体 (Mab) 或抗体碎片抗原结合 (Fab) 区域在中国仓鼠卵巢 (CHO) 细胞中.
在表达后,抗体:抗原复合物被净化,使用低pH处理与氨酸分离,并通过制剂尺寸排除染色学 (pSEC) 分离单个成分,可选的亲和染色学用于残留伴侣去除.

主要成果:

在传统方法 (histidine-tagging, Fc-fusion) 失败的情况下,成功地为来自分化/细胞因子家族的两个具有挑战性的目标产生了稳定和生物活性的ECD抗原.
证明了抗体 - 沙佩龙介导的联合表达和解离方法的有效性,用于产生难以表达的抗原.

结论:

抗体-沙佩龙介导的联合表达/解离方法为产生稳定,功能细胞外域 (ECD) 抗原提供了强大的和可泛化的方法.
这种技术为克服治疗性抗体开发至关重要的抗原的生产挑战提供了可行的解决方案.

关键词:

这就是BTLA.这就是为什么Fab Fab是Fab.在IMAC中,IMAC是IMAC.有的可能是 Mab Mab Mab Mab.美国证券交易委员会 SEC SEC.活动测定试验.抗体:是一种抗原复合体.抗原ECD是一种抗原ECD.在阿根廷,阿根廷人.陪伴者是一个陪伴者.酸是一种酸.共同表达的共同表达.细胞因子细胞因子分离缓冲区分离缓冲区分

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