Quality control of recombinant adeno-associated virus type 2/human blood coagulation factor IX.
- Author:
Kai GAO
1
;
Jun-zhi WANG
;
Chun-ming RAO
;
Xiao-bing WU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Dependovirus; genetics; Factor IX; biosynthesis; genetics; Female; Gene Transfer Techniques; Genetic Vectors; genetics; Genome, Viral; Humans; Male; Mice; Mice, Knockout; Quality Control; Random Allocation; Recombinant Proteins; biosynthesis; genetics
- From: Acta Pharmaceutica Sinica 2003;38(9):684-689
- CountryChina
- Language:Chinese
-
Abstract:
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 x 10(15) VG.L-1. hFIX expression was more than 20.0 micrograms.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.
