Department of Biomedical Engineering, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
Department of Radiology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
PLoS One. 2018 Oct 23;13(10):e0206246. doi: 10.1371/journal.pone.0206246. eCollection 2018.
Antibody-drug conjugate (ADC) is a class of targeted cancer therapies that combine the advantages of monoclonal antibody (mAb)'s specific targeting and chemotherapy's potent cytotoxicity. The therapeutic effect of ADC is significantly affected by its bioproduction process. This study aims to develop an effective ADC production process using anti-HER2 mAb-drug as a model therapeutic. First, a high titer (>2 g/L) of mAb was produced by Chinese hamster ovary cells from fed-batch cell culture. Both live-cell confocal microscopy imaging and flow cytometry analysis demonstrated that the produced mAb and ADC had strong and specific binding to HER2+ cell line BT474. Second, various conjugation conditions of mAb and drug, including linker selection, ratio of drug and mAb, and conjugation approaches, were investigated to improve the production yield and product quality. Finally, the ADC structure and biological quality were evaluated by SDS-PAGE and anti-breast cancer toxicity study, respectively. The ADC with integral molecular structure and high cytotoxicity (IC50 of 1.95 nM) was produced using the optimized production process. The robust bioproduction process could guide the development of ADC-based biopharmaceuticals.
抗体偶联药物（ADC）是一类靶向癌症治疗药物，结合了单克隆抗体（mAb）的特异性靶向和化疗的强大细胞毒性的优势。ADC 的治疗效果受到其生物生产工艺的显著影响。本研究旨在使用抗 HER2 mAb-药物作为模型治疗药物，开发一种有效的 ADC 生产工艺。首先，通过灌流细胞培养从中国仓鼠卵巢细胞中生产出高滴度（>2g/L）的 mAb。活细胞共聚焦显微镜成像和流式细胞术分析表明，所产生的 mAb 和 ADC 与 HER2+细胞系 BT474 具有强烈和特异性结合。其次，研究了 mAb 和药物的各种偶联条件，包括连接子的选择、药物与 mAb 的比例以及偶联方法，以提高生产产率和产品质量。最后，通过 SDS-PAGE 和抗乳腺癌毒性研究分别评估了 ADC 的结构和生物学质量。使用优化的生产工艺生产出具有完整分子结构和高细胞毒性（IC50 为 1.95 nM）的 ADC。稳健的生物生产工艺可以为基于 ADC 的生物制药的开发提供指导。
