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基于细胞的蛋白质阵列的出现,用于测试抗体治疗药物的多特异性目标外结合
Diana M Norden1, Carmen T Navia1, Jonathan T Sullivan1
1Integral Molecular, Philadelphia, PA, USA.
mAbs
|August 24, 2024
概括
与传统的组织交叉反应性研究相比,基于细胞的蛋白质阵列为评估单克隆抗体 (mAb) 特异性提供了更可靠的方法. 这种改进的特异性分析可以减少目标外结合,可能防止生物治疗开发中的不良事件.
科学领域:
- 生物化学 生物化学
- 免疫学 免疫学 免疫学
- 药物开发 药物开发
背景情况:
- 在人类试验之前,对单克隆抗体 (mAbs) 和CAR-T细胞的特异性分析至关重要.
- 对于mAb特异性的传统组织交叉反应性研究是不可靠的,往往缺少目标外结合.
- 目标外结合可能导致生物治疗开发中未被检测到的问题.
研究的目的:
- 审查基于细胞的蛋白质阵列作为评估mAb特异性的改进方法.
- 突出传统特异性评估方法的局限性.
- 提出关于治疗抗体中脱结合的流行情况的数据.
主要方法:
- 对基于细胞的蛋白质阵列技术进行特异性评估的审查.
- 对全人类膜蛋白质组 (约6,000种蛋白质) 结合的分析.
- 膜蛋白在活细胞或未固定细胞内的本地配置中的表达.
主要成果:
- 基于细胞的蛋白质阵列提供了对mAb特异性的全面评估.
- 观察到超标结合率高:33%在候选药物中,20%在销售的治疗药物中.
- 目标外结合被认为是不良事件和药物消耗的重要原因.
结论:
- 基于细胞的蛋白质阵列代表了mAb特异性分析的优越替代方案.
- 在发育早期解决非目标结合对于生物治疗安全性和成功至关重要.
- 改进的特异性评估可以减轻与治疗抗体相关的风险.
相关概念视频
00:42Cross-reactivity
Overview
01:15Western Blotting
Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
01:33Enzyme-Linked Immunosorbent Assay
In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.