[Quality control of clinical-grade recombinant adenovirus used in gene therapy].
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.
Zhang
Xiao-zhi
XZ
Shenzhen SiBiono GeneTech Co. Ltd, Shenzhen 518057, China.
Lin
Hong
H
Yang
Xiao-yan
XY
Wang
Cai-hong
CH
Zhang
Ming-ming
MM
Chen
Pei-la
PL
Peng
Zhao-hui
ZH
D000256:Adenoviridae / Q000235:genetics*; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D015316:Genetic Therapy / Q000379:methods* / Q000592:standards; D005822:Genetic Vectors / Q000235:genetics / Q000592:standards; D006801:Humans; D011786:Quality Control; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D016159:Tumor Suppressor Protein p53