Jeong Yun Hee, Lennon Gillian, Veldman Geertruida, Serna Daniel M, Ibrahimov Alexander
In Vitro Immunosafety, Development Biological Sciences, Abbvie Bioresearch Center, Worcester, MA, USA.
Biotherapeutics Discovery Research, AbbVie Bioresearch Center, AbbVie Bioresearch Center, Worcester, MA, USA.
MAbs. 2025 Dec;17(1):2458627. doi: 10.1080/19420862.2025.2458627. Epub 2025 Feb 1.
Immunogenic responses to biotherapeutics often lead to termination of their development because the resulting anti-drug-antibodies (ADA) can negatively impact pharmacology, safety, and efficacy. To mitigate ADA risks, in vitro risk assessment assays in non-clinical settings are essential to enhance safety and efficacy of protein-based therapeutics. This study aimed to develop and validate a human immunogenicity T cell proliferation assay. However, there is a lack of comprehensive guidelines for managing product-related factors such as endotoxin contamination, which can significantly influence assay sensitivity and accuracy. Our investigation of the impact of endotoxins revealed that levels above 0.1 EU/mg significantly induce T cell proliferation and CD14 myeloid cell expansion, leading to potential false-positive outcomes in immunogenicity assessments. These findings suggest the importance of developing standardized protocols to enhance the predictive capability of in vitro methods, ensuring the assessment of therapeutic proteins accurately reflects their immunogenic potential without interference from contaminants.
对生物疗法的免疫原性反应常常导致其研发终止，因为产生的抗药抗体（ADA）会对药理学、安全性和疗效产生负面影响。为降低ADA风险，非临床环境中的体外风险评估测定对于提高基于蛋白质的治疗药物的安全性和疗效至关重要。本研究旨在开发并验证一种人免疫原性T细胞增殖测定法。然而，在管理诸如内毒素污染等与产品相关的因素方面缺乏全面的指导方针，而这些因素会显著影响测定的灵敏度和准确性。我们对内毒素影响的调查显示，高于0.1 EU/mg的水平会显著诱导T细胞增殖和CD14髓样细胞扩增，从而导致免疫原性评估中出现潜在的假阳性结果。这些发现表明制定标准化方案以提高体外方法预测能力的重要性，确保治疗性蛋白质的评估能准确反映其免疫原性潜力，而不受污染物干扰。
