Abstract：
In the present study the compositional analysis of the amino acids released by the acidic hydrolysis of the vaccine antigens was approached as an alternative to the dye-binding methods, for improvement of the quality control. In particular, the Analytical Quality by Design principles were undertaken in optimizing the hydrolysis conditions of the antigens to be applied prior to the quantitation by UHPLC-UV. Bexsero was used as a case study; it is a recombinant meningococcal B vaccine and one of its critical quality attributes is the content of the three core protein antigens, namely Neisseria Heparin Binding Antigen, factor H binding protein and Neisseria adhesin A, in the final formulation. Conventionally, the proteins quantitation is carried out by dye-binding assays. Analytical Target Profile was defined as the accurate determination of amounts of the Bexsero antigens. The Critical Method Parameters were chosen by means of the cause-effect matrix. A Face Centered Design was used to select the experiments to investigate the process and finally a Method Operable Design Region with a risk of failure of 5% was defined. The selected working point for routine use was: hydrolysis time, 17 hrs; temperature, 112 °C; 6 M HCl volume, 300 µl; antioxidant 90% phenol volume, 5 µl.
摘要：
在本研究中，通过疫苗抗原的酸性水解释放的氨基酸的组成分析被作为染料结合方法的替代方法。用于质量控制的改进。特别是,在通过UHPLC-UV定量之前,在优化待应用的抗原的水解条件中进行设计原理的分析质量。Bexsero被用作案例研究；它是一种重组脑膜炎球菌B疫苗，其关键质量属性之一是三种核心蛋白抗原的含量，即奈瑟菌肝素结合抗原，H因子结合蛋白和奈瑟菌粘附素A,在最后的配方中。传统上,蛋白质定量是通过染料结合测定法进行的。分析目标曲线定义为Bexsero抗原的量的准确测定。通过因果矩阵选择关键方法参数。使用以面为中心的设计来选择实验以研究过程，最后定义了失败风险为5％的方法可操作设计区域。常规使用的选定工作点是：水解时间，17小时；温度，112°C;6MHCl体积,300μl;抗氧化剂90%苯酚体积,5µl
