中国药理学通报2016，Vol.32Issue(6)：881-884,885,5.DOI:10.3969/j.issn.1001-1978.2016.06.027
人AS PP2重组腺病毒质量控制研究
Quality control of human ASPP2 recombinant adenovirus
摘要
Abstract
Aim Toanalyzethekeyqualityandestablishmeth-ods for essential quality control of human recombinant ASPP2 adenovirus.Methods TheviralstructuralgeneofE2Bandtar-get gene of ASPP2 were identified by PCR;The number of virus particles was measured by UV-SDS methods;Infectious titer was determined by TCID50 assay;Target protein of ASPP2 was ob-served by Western blot assay;The biological effects of recombi-nant adenovirus on liver cancer cells were evaluated by MTT as-say;A549 cells were used to check replication of the competent adenovirus(RCA)by the observation of the cytopathic effect. Results PCRanalysisofE2BandASPP2wasinconsistent with theoretical values;Particle numbers of virus were 5. 6 × 1012 VP/mL,infectious titer was 2 ×1011 IU/mL and specific activity was 3. 5%;ASPP2 protein expression could be detected when cells were infected with virus for 24 h;Growth inhibition of liver cancer cells could be found by adding recombinant ASPP 2 adenovirus;The level of RCA was less than 1 RCA/3. 0 ×1010 VP,in line with the standards of China Food and Drug Adminis-tration(SFDA).Conclusion Thequalitycontrolmethodswere established aiming at key characters of human recombinant AS-PP2 adenovirus,which may provide foundations for its quality standard and future applications.关键词
基因治疗/腺病毒/ASPP2/质量控制/病毒比活/MTTKey words
gene therapy/adenovirus/ASPP2/quality control/virus specific activity/MTT分类
医药卫生引用本文复制引用
王爽,许坚吉,刘晓霓,陈德喜..人AS PP2重组腺病毒质量控制研究[J].中国药理学通报,2016,32(6):881-884,885,5.基金项目
国家自然科学基金资助项目（No 81272266）；北京市卫生系统高层次卫生技术人才培养项目资助（No 2015-3-101）；国家自然科学基金国际（地区）合作与交流项目（No 81361120401）；北京市肝病研究所自主课题基金 （）
