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在抗体CH2域中的L309K突变的药物适应性特性

Cui Li¹, Qing-Qing Yao², Jiang Li¹

¹Department of Pharmacy, Zhejiang Provincial Hospital of Chinese Medicine, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, 310000 Zhejiang China.

|May 14, 2024

概括

通过减少聚合和改善物理化学性质,L309K突变增强了抗体的药物可用性. 这种抗体工程策略维持生物功能,同时提高稳定性和减少不必要的Fc受体相互作用.

科学领域:

生物制药的发展.
蛋白质工程是一种蛋白质工程.
免疫学免疫学免疫学

背景情况:

抗体药物开发需要通过物理化学性质评估优化药物可用性.
表面暴露的疏水区域可能会导致抗体聚合和杂质形成.
候选抗体aPDL1-WT在早期发展阶段提出了挑战.

研究的目的:

评估L309K突变策略,以提高aPDL1-WT抗体的药性.
评估L309K突变对物理化学性质和压力条件下的聚合的影响.
确定L309K突变对生物功能和Fc受体结合的影响.

主要方法:

部位定向突变发生引入aPDL1-WT中的L309K突变,产生aPDL1-LK.
加快高温稳定性研究,以评估聚合和杂质形成.
物理化学性质的表征,包括水友性,形状稳定性和电荷变异.
对hPDL1,Fc受体 (CD16a,CD64,CD32a) 的结合亲和度测定,并补充C1q.
在体外功能测定:抗体依赖的细胞介导细胞毒性 (ADCC),抗体依赖的细胞化 (ADCP) 和补体依赖的细胞毒性 (CDC).

主要成果:

与aPDL1-WT相比,aPDL1-LK变异在加速高温条件下显著减少聚合和大分子杂质.
aPDL1-LK表现出改善的物理化学特性,包括增强的水友性和稳定性.
L309K突变并没有改变glycosylation或对hPDL1和几个Fc受体/补充成分的结合亲和力.
对于aPDL1-LK.K.观察到对CD16a-V158和CD32a-R131的亲和力减少了两倍.
L309K突变可能会降低CD16a-V158介导的ADCC活性,但不会影响CD32a-R131介导的ADCP活性.

结论:

L309K突变是一种可行的策略,可以增强抗体候选者的药用性.
这种突变提高了抗体的稳定性,并减少了聚合,而不会影响抗原结合或基本的生物功能.
通过L309K调节Fc受体结合亲和力,为微调抗体效应器功能提供了一种途径.

关键词:

抗体是对抗体的一种.药物适应性属性药物适应性属性突变突变是一种突变.表面暴露的疏水性残留物.

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