[Quality control for recombinant adenovirus-IL2].
OBJECTIVE
To investigate the quality and optimized test methods and establish the quality specification of recombinant adenovirus-IL2.


METHODS
The titer of Adv-IL2 was measured by cytopathic effect (CPE). Hela cells were infected with Adv-IL2 in vitro, the expressed IL-2 and its bioactivity in Hela cell were determined by enzyme-linked immunosorbent assay (ELISA) and 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) respectively. The purity of Adv-IL2 was analysed by UV and IE-HPLC method. The molecular weight and enzyme digestive map of Adv-IL2 genome were analysed by electrophoresis. The characteristic gene E2B, IL-2 expression casseter and foreign factors (RCV, HIV, HBV, HCV) were detected with polymerase chain reaction (PCR). Other tests were carried out according to the Chinese Requirements for Biological Products.


RESULTS
Adv-IL2 was generated efficiently with a titer of 3 x 10(10) pfu.mL-1. The expressed IL-2 and its bioactivity were 25 ng.mL-1 and 700 u.mL-1 respectively. A260 nm/A280 nm was 1.23. The purity determined by IE-HPLC was higher than 98%. The molecular weight, enzyme digestive map of Adv-IL2 genome, the characteristic gene E2B and IL-2 expression casseter conformed to expected values.


CONCLUSIONS
The specification for Adv-IL2 is established and can be used for the quality control of the product.
Lin
Jian-wei
JW
National Institute for the Control of Pharmaceutical and Biological Products, Beijing 100050, China.
Wang
Jun-zhi
JZ
Rao
Chun-ming
CM
D000256:Adenoviridae / Q000235:genetics*; D002460:Cell Line; D004622:Embryo, Mammalian; D005822:Genetic Vectors / Q000235:genetics; D006367:HeLa Cells / Q000821:virology; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis* / Q000235:genetics; D011786:Quality Control; D011994:Recombinant Proteins / Q000096:biosynthesis; D014162:Transfection
D007376:Interleukin-2; D011994:Recombinant Proteins