发布时间 : 星期五 文章WHO对HPV疫苗质量、安全性及有效性指导原则更新完毕开始阅读cd5f5a1f0c22590103029d39
monovalent antigen bulk adsorbed on an aluminium containing adjuvant. Different batches of adsorbed monovalent antigen bulks may be pooled before collection into a single vessel.
Adjuvant: A vaccine adjuvant is a component that potentiates the immune response to an antigen and/or modulates it towards the desired immune responses.
Final vaccine bulk: The formulated bulk present in the container from which the final containers are filled. The final bulk may be prepared from one or more adsorbed monovalent antigen bulks and may contain VLP antigens from one or multiple HPV virus types.
Filling lot (final vaccine lot): A collection of sealed final containers of vaccine that is homogeneous with respect to the risk of contamination during the filling process. A filling lot must therefore have been filled or prepared in one working session. 3.3生产建议
生产必须符合GMP要求，生物安全上要求无菌。不同类型HPV L1 VLP应分别生产，同时，还要有充分的清洁验证。
抗原生产过程，须验证以证明生产的稳定性，至少要连续三批。
但如两种蛋白的纯化步骤一样，可只用验证一种。生产一致性评估应包括关键质量参数评估及它们相应的特征，如宿主DNA与HCP清除、工艺过程系数如柱负荷。不同批次抗原蛋白生产过程验证应与之前
HPV VLP生产的规格保持一致如抗原特性和纯度。 3.3.1鉴定
HPV的鉴定应进行多批次疫苗生产中进行，包括过程验证中的批次。 蛋白质组成验证：还原SDS－PAGE，并对条带进行染色，如有可能最好用相应抗体或质谱技术，确认L1目的蛋白存在。 蛋白完整性（identity）验证：肽谱图和末端氨基酸序列分析。 空间表位对有效性有着重大的影响，因而必须检测分析VLP的形态特征及聚合程度。此外，在合适情况下，应检测蛋白质，脂质、核酸和碳水化合物等。
VLP特征参数可通过下面这些技术获得：原子力和透射电子显微镜、动态散光、表位作图、单抗中和反应。宿主蛋白质残留应满足非临床和临床要求（参见Part B and C） 3.3原材料控制
3.3.1 抗原生产细胞培养物 Cell cultures for antigen production The use of any cell line should be based on a cell bank system. Only cells that have been approved and registered with the national regulatory authority should be used to produce HPV L1 protein. The national regulatory authority should be responsible for approving the cell bank. Appropriate history of the cell bank should be provided. 3.3.1.1 Yeast cells
The characteristics of the recombinant production strain (host cell in combination with the expression vector system) should be fully described
and information given on the absence of adventitious agents and on gene homogeneity for the master and working cell banks. A full description of the biological characteristics of the host cell and expression vectors should be given. The physiological measures used to promote and control the expression of the cloned gene in the host cell should be described in detail. This should include genetic markers of the host cell, the construction, genetics and structure of the expression vector and the origin and identification of the gene that is being cloned.
The nucleotide sequence of the gene insert and of adjacent segments of the vector and restriction?enzyme mapping of the vector containing the gene insert should be provided as required by the national control authority. 其他略
3.3.2 细胞培养基 Cell culture medium
如添加血清，应验证无抗生素。略
3.3.3主细胞库和工作细胞验证 Tests on master and working cell banks 略
3.4 HPV疫苗生产控制
3.4.1酵母表达体系中单一表达HPV抗原生产控制 3.4.1.1微生物纯度 略 略
3.5纯化后单价抗原质量控制 3.5.1 鉴定：免疫学测定
3.3.2 纯度：还原SDS-PAGE，考染染色后，对条带进行灰度计算，分析目的条带纯度
3.3.3蛋白含量：总蛋白的量可用凯氏定氮法，或Lowry法，及Brandford
3.3.4 抗原含量：用专一性强的方法，对纯化各环节的抗原进行定量分析。此外，在纯化环节中，还应对抗原含量与总蛋白质的比值进行计算。 3.3.5 无菌检查 3.3.6 L1单体完整率
生产稳定后，采用合适的方法（什么方法？）对L1单体完整率进行测定
3.3.7 VLP大小和结构：电镜等 3.3.8工艺相关杂质测定
在生产中，所使用的任何有潜在负作用的试剂，都应对其残留
进行检测。在工艺稳定后，可以不必进行日常检测。
宿主DNA与宿主蛋白残留检测，方法应为专一性好、灵敏度
高的分析方法。DNA残留水平不得超过规定水平。工艺稳定后，可以不必日常检测。
3.3.9 血清蛋白含量与病毒清除 略