Quality control of recombinant adeno-associated virus type 2/human blood coagulation factor IX

AimTo establish quality control requirements and methods for recombinant adeno-associated virus（rAAV）type 2/human blood coagulation factor IX (rAAV-2/hFIX). MethodsIdentification of rAAV genome fragments, potential contaminants including wild type AAV(wtAAV) and helper virus, were detected by PCR. Purity of rAAV-2/hFIX was analyzed by cation-exchange HPLC and SDS-PAGE. Virus partical numbers were performed by dot blot assay. hFIX expression was demonstrated by ELISA and potency of hFIX was verified by APTT. ResultsIdentity of rAAV-2/hFIX was proved. Residues of wtAAV and helper virus were conformed to requirements. Purity of rAAV-2/hFIX were more than 98%. Partical numbers of rAAV-2/hFIX were more than 1.0×10₁₅VG·L^-1. hFIX expression was more than 20.0 μg·L^-1. hFIX potency was verified by APTT following rAAV-2/hFIX injected to FIX gene knockout mice, potency results conformed to requirements. ConclusionThe methods and requirements had been established for quality control of rAAV-2/hFIX.