Quality control of recombinant adeno-associated virus type 2/human blood coagulation factor IX / 药学学报
Acta Pharmaceutica Sinica
;
(12): 684-689, 2003.
Article
in Chinese
| WPRIM
| ID: wpr-266589
ABSTRACT
<p><b>AIM</b>To establish quality control requirements and methods for recombinant adeno-associated virus(rAAV) type 2/human blood coagulation factor IX (rAAV-2/hFIX).</p><p><b>METHODS</b>Identification 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.</p><p><b>RESULTS</b>Identity 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.</p><p><b>CONCLUSION</b>The methods and requirements had been established for quality control of rAAV-2/hFIX.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Quality Control
/
Recombinant Proteins
/
Factor IX
/
Random Allocation
/
Genome, Viral
/
Mice, Knockout
/
Gene Transfer Techniques
/
Dependovirus
/
Genetic Vectors
/
Genetics
Type of study:
Controlled clinical trial
/
Prognostic study
Limits:
Animals
/
Female
/
Humans
/
Male
Language:
Chinese
Journal:
Acta Pharmaceutica Sinica
Year:
2003
Type:
Article
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