摘要
目的 建立并完善我国重组腺病毒临床级基因治疗制品重组人 p5 3腺病毒注射液的质量控制标准和方法。方法 采用组织培养半数感染剂量 (TCID50 )方法测定病毒的感染滴度 ;采用HPLC进行病毒的纯度分析 ;采用A5 4 9细胞进行样品的复制型病毒检测 ,其他质量控制方法按照中国生物制品规程进行。结果 重组人p5 3腺病毒注射液 (Lot 2 0 0 10 70 1)成品的病毒颗粒数为 1 0 3×10 12 VP/ml,病毒滴度为 5 0 1× 10 10 IU/ml;比活性为 4 86 % ,纯度达到 98 6 2 % ,复制型腺病毒(RCA)水平小于 1RCA/ 3× 10 10 VP ,符合中国食品药品监督管理局 (SFDA)的标准。结论 建立了重组腺病毒临床级基因治疗制品的全套质量标准 ,达到SFDA的质量要求 ,可有效地控制制品的质量 ,保证重组腺病毒制品在临床应用中安全有效。
Objective To establish the quality control methods and reference standards for clinical-grade recombinant adenovirus products for human gene therapy. Methods The Infectivity of clinical grade recombinant adenoviral vectors is determined by a TCID 50 assay. The purity is determined by a high-performance liquid chromatography (HPLC) assay. A549 cells were used in replication competent adenovirus (RCA) assay of samples by observation of the cytopathic effect. Other quality control assays were performed in accordance with the SFDA Regulations for Biological Products. Results The recombinant adenovirus encoding human p53 gene produced at SiBiono(Lot 20010701)has the following quality attributes: viral particle concentration: 1.03(10 12 VP/ml,infectivity titer: 5.01(10 10 IU/ml,specific infectivity (IU/VP): 4.86%,higher than the 3.3% required by the Food and Drug Administration (FDA),USA; Purity by HPLC analysis: 98.62%,higher than the 95% purity specified by SFDA; and level of RCA: less than 1 RCA / 3(10 10 VP,meeting the standards established by SFDA. Conclusions A whole set of quality standards of clinical-grade recombinant adenovirus vectors has been established so as to ensure the clinical safety and efficacy of recombinant adenoviral vectors for human gene therapy.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2004年第10期849-852,共4页
National Medical Journal of China
基金
国家"十五"重大科技专项创新药物和中药现代化基金资助项目 (2 0 0 2AA2Z3 3 0 1)
