Qin Dan, Phung Qui, Wu Patrick, Yin Zhaojun, Tam Sien, Tran Peter, ElSohly Adel M, Gober Joshua, Hu Zicheng, Zhou Zhenru, Cohen Sivan, He Dongping, Bainbridge Travis W, Kemball Christopher C, Zarzar Jonathan, Sreedhara Alavattam, Stephens Nicole, Decalf Jérémie, Moussion Christine, Ye Zhengmao, Balazs Mercedesz, Li Yinyin
Biochemical and Cellular Pharmacology, Genentech Inc., South San Francisco, CA, United States.
Proteomic and Genomic Technologies, Genentech Inc., South San Francisco, CA, United States.
Front Immunol. 2025 Feb 13;16:1519452. doi: 10.3389/fimmu.2025.1519452. eCollection 2025.
As a crucial asset for human health and modern medicine, an increasing number of biotherapeutics are entering the clinic. However, due to their complexity, these drugs have a higher potential to be immunogenic, leading to the generation of anti-drug antibodies (ADAs). Clinically significant ADAs have an impact on pharmacokinetics (PK), pharmacodynamics (PD), effectiveness, and/or safety. Thus, it is crucial to understand, manage and minimize the immunogenicity potential during drug development, ideally starting from the molecule design stage.
In this study, we utilized various immunogenicity risk assessment methods, including prediction, dendritic cell internalization, MHC-associated peptide proteomics, HLA peptide binding, and T cell proliferation, to assess the immunogenicity risk of FLT3L-Fc variants.
We identified a single point mutation in the human FLT3L-Fc protein that introduced highly immunogenic T cell epitopes, leading to the induction of T cell responses and thereby increasing the immunogenicity risk in clinical settings. Consequently, the variant with this point mutation was removed from further consideration as a clinical candidate.
This finding underscores the necessity for careful evaluation of mutations during the engineering of protein therapeutics. The integration of multiple immunogenicity risk assessment tools offers critical insights for informed decision-making in candidate sequence design and therapeutic lead selection.
作为人类健康和现代医学的关键资产，越来越多的生物治疗药物进入临床。然而，由于其复杂性，这些药物具有更高的免疫原性潜力，导致抗药物抗体（ADA）的产生。具有临床意义的ADA会对药代动力学（PK）、药效学（PD）、有效性和/或安全性产生影响。因此，在药物开发过程中，理想情况下从分子设计阶段开始，了解、管理并最小化免疫原性潜力至关重要。
在本研究中，我们利用了各种免疫原性风险评估方法，包括预测、树突状细胞内化、MHC相关肽蛋白质组学、HLA肽结合和T细胞增殖，以评估FLT3L-Fc变体的免疫原性风险。
我们在人FLT3L-Fc蛋白中鉴定出一个单点突变，该突变引入了高度免疫原性的T细胞表位，导致T细胞反应的诱导，从而增加了临床环境中的免疫原性风险。因此，具有该点突变的变体被排除在作为临床候选药物的进一步考虑之外。
这一发现强调了在蛋白质治疗药物工程过程中仔细评估突变的必要性。多种免疫原性风险评估工具的整合为候选序列设计和治疗先导物选择中的明智决策提供了关键见解。
