Quality control of human ASPP2 recombinant adenovirus
10.3969/j.issn.1001-1978.2016.06.027
- VernacularTitle:人AS PP2重组腺病毒质量控制研究
- Author:
Shuang WANG
;
Jianji XU
;
Xiaoni LIU
;
Dexi CHEN
- Publication Type:Journal Article
- Keywords:
gene therapy;
adenovirus;
ASPP2;
quality control;
virus specific activity;
MTT
- From:
Chinese Pharmacological Bulletin
2016;32(6):881-884,885
- CountryChina
- Language:Chinese
-
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.
