Quality control for recombinant adenovirus-IL2 / 药学学报

ABSTRACT

<p><b>AIM</b>To investigate the quality and optimized test methods and establish the quality specification of recombinant adenovirus-IL2.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>The specification for Adv-IL2 is established and can be used for the quality control of the product.</p>