Ajinomoto Co., Inc., 1-1 Suzuki-cho, Kawasaki, Kawasaki, Kanagawa, 210-8681, Japan.
Ajinomoto Bio-Pharma Services, 11040 Roselle Street, San Diego, CA, 92121, United States.
Anal Sci. 2021 Aug 10;37(8):1171-1176. doi: 10.2116/analsci.20P424. Epub 2021 Jan 29.
The field of oncology has recently seen an exponential growth in antibody-drug conjugates (ADCs) as a biopharmaceutical class with seven ADCs being launched onto the market in the last ten years. Despite the increase in the industrial research and development of these compounds, their structural complexity and heterogeneity continue to present various challenges regarding their analysis including reaction monitoring. Robust and simple reaction monitoring analysis are in demand in the view of at-line in-process monitoring, and can instill control, confidence and reliability in the ADC manufacturing process. Aiming at providing chromatographic methods for conjugation monitoring, we evaluated herein the potential of utilizing reverse phase HPLC analysis, without sample pretreatment, for characterization of traditional cysteine-based ADCs. This analysis can be used for estimation of drug antibody ratio (DAR), which has shown the same trends and results as other well-established HPLC techniques. This methodology was also applied to three ADCs derived from three different antibodies. Additionally, we analyzed unpurified ADC samples existing in a complex reaction matrix and separated ADC species and payload compounds. This investigation was conducted using three different ADCs based on different payloads. The results described herein indicate the potential application of this RP-HPLC methodology in reaction monitoring studies.
近年来，抗体药物偶联物（ADC）作为一种生物制药类别，在肿瘤学领域呈指数级增长，在过去十年中已有七种 ADC 上市。尽管这些化合物的工业研发有所增加，但它们的结构复杂性和异质性仍然对其分析提出了各种挑战，包括反应监测。鉴于在线过程监测的需求，对强大且简单的反应监测分析的需求不断增加，这可以为 ADC 制造过程提供控制、信心和可靠性。本研究旨在为缀合监测提供色谱方法，评估了在此无需样品预处理的情况下，反相 HPLC 分析在传统半胱氨酸 ADC 表征中的应用潜力。该分析可用于估计药物抗体比（DAR），其与其他成熟的 HPLC 技术显示出相同的趋势和结果。该方法还应用于三种源自三种不同抗体的 ADC。此外，我们分析了存在于复杂反应基质中的未纯化 ADC 样品，并分离了 ADC 种类和有效载荷化合物。本研究使用三种基于不同有效载荷的不同 ADC 进行。本文所述的结果表明，该反相 HPLC 方法在反应监测研究中具有潜在的应用。
