Houy Camille, Ming Marin, Ettorre Luciano, Jin Robbie, Thangavadivel Nemika, Chen Tricia, Su Jin, Gajewska Beata
Department of Analytical Sciences, Sanofi Pasteur, Toronto, ON M2R 3T4, Canada.
Vaccines (Basel). 2022 May 13;10(5):775. doi: 10.3390/vaccines10050775.
In the vaccine industry, multiple physicochemical, immunological, and analytical methods are applied throughout the manufacturing process to characterize and monitor the quality of vaccines. Presented here is the Single Epitope Antigenicity Test (SEAT), an innovative, quantitative epitope profiling method which provides an extended immunochemical analysis for diphtheria toxoid (DTxd) to be used for consistency testing during manufacturing process changes. The method uses BioLayer Interferometry (BLI) and a panel of monoclonal antibodies (mAbs) to independently assess nine individual antigenic sites of DTxd. The panel includes mAbs which are functional, bind distinct sites on DTxd and are able to distinguish intact DTxd from that which has been exposed to heat treatment. The SEAT method was qualified for precision, accuracy, and linearity, and was used to define a preliminary comparability range for DTxd made using the current manufacturing process. DTxd lots manufactured using alternate processes were assessed in the context of this range to determine the impact on DTxd antigenicity. Epitope profiling by SEAT provides quantitative information on the integrity of multiple important antigenic regions of DTxd, and therefore represents a valuable tool in a comprehensive analytical test package which can be used to support manufacturing process changes for vaccines.
在疫苗行业，整个生产过程中会应用多种物理化学、免疫学和分析方法来表征和监测疫苗质量。本文介绍了单表位抗原性测试（SEAT），这是一种创新的定量表位分析方法，可为白喉类毒素（DTxd）提供扩展的免疫化学分析，用于生产工艺变更期间的一致性测试。该方法使用生物层干涉术（BLI）和一组单克隆抗体（mAb）来独立评估DTxd的九个单独抗原位点。该组包括具有功能的mAb，它们结合DTxd上不同的位点，并且能够区分完整的DTxd和经过热处理的DTxd。SEAT方法在精密度、准确性和线性方面得到了验证，并用于确定使用当前生产工艺制备的DTxd的初步可比性范围。在此范围内评估使用替代工艺生产的DTxd批次，以确定对DTxd抗原性的影响。通过SEAT进行的表位分析提供了关于DTxd多个重要抗原区域完整性的定量信息，因此是综合分析测试包中的一个有价值的工具，可用于支持疫苗生产工艺的变更。
