Department of Molecular Systems BioAnalysis, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8501, Japan; Analytical and Quality Evaluation Research Laboratories, Daiichi Sankyo Co., Ltd., Hiratsuka, Kanagawa 254-0014, Japan.
Analytical and Quality Evaluation Research Laboratories, Daiichi Sankyo Co., Ltd., Hiratsuka, Kanagawa 254-0014, Japan.
J Chromatogr A. 2022 Dec 6;1685:463645. doi: 10.1016/j.chroma.2022.463645. Epub 2022 Nov 11.
We evaluated the immunoreactivity profiles of eight commercial anti-host cell protein (anti-HCP) antibodies from different host animals and their antigens used for immunization by an isobaric labeled affinity purification-mass spectrometry (AP-MS) method. As a result, 34 proteins with high abundance but low recovery from harvest cell culture fluid were identified. Since they are likely to be underestimated in biopharmaceutical quality assessment, the features common to these proteins were investigated. Compared to other immunoprecipitated HCP proteins, proteins exhibiting lower molecular weight (ΔMW = -14600), lower isoelectric point (ΔpI = -0.86), and lower hydrophobicity (ΔGRAVY = -0.13) were enriched. This AP-MS method provides important information for HCP control strategies using immunological methods and is expected to contribute to the development of safe biopharmaceutics.
我们采用等压标记亲和纯化-质谱（AP-MS）方法评估了来自不同宿主动物的八种商业化抗宿主细胞蛋白（anti-HCP）抗体及其免疫原的免疫反应性谱。结果，鉴定出 34 种高丰度但从收获细胞培养液中回收率低的蛋白质。由于它们在生物制药质量评估中可能被低估，因此研究了这些蛋白质的共同特征。与其他免疫沉淀的 HCP 蛋白相比，富集了分子量较低（ΔMW=-14600）、等电点较低（ΔpI=-0.86）和疏水性较低（ΔGRAVY=-0.13）的蛋白质。该 AP-MS 方法为使用免疫学方法进行 HCP 控制策略提供了重要信息，有望为安全的生物制药的发展做出贡献。
