Packaging of rAAV-Endostatin and expressing in EJ cells

Objective:To package rAAV-Endostatin for endostatin gene therapy of bladder cancer.Methods:To construct pAAV-IgG-Endostatin and identified by enzyme digesting and sequencing method.rAAV-Endostatin was packaged by co-transfection technique then identified under electronic microscope and assayed by ELISA.EJ cells were transfected by rAAV-Endostatin and endostatin concentration in supernatant was assayed by ELISA too.Results:pAAV-IgG-Endostatin was constructed and rAAV-Endostatin was packaged successfully,virus titer is 1.0×1012 v.p/ml,virus pattern is normal.After EJ cells being transfected,endostatin was expressed and secreted,endostatin concentration in the supernatant is 54.09 ng/ml.Conclusion:rAAV-Endostatin was packaged successfully and thus endostatin gene therapy of bladder cancer was made possible.