QUALITY CONTROL FOR RECOMBINANT ADENOVIRUS-IL2

AIMTo investigate the quality and optimized test methods and establish the quality specification of recombinant adenovirus-IL2. METHODSThe 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. RESULTSAdv-IL2 was generated efficiently with a titer of 3×10¹⁰ pfu·mL^-1. The expressed IL-2 and its bioactivity were 25 ng·mL^-1 and 700 u·mL^-1 respectively. A_{260 nm}/A_{280 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. CONCLUSIONThe specification for Adv-IL2 is established and can be used for the quality control of the product.