[Characterization of the biological activities of lyophilized recombinant adenovirus expressing the triple mutant of hypoxia inducible factor-1α].

Abstract

OBJECTIVE:

To investigate the influence of lyophilization on the biological activity of recombinant adenovirus-mediated triple mutant of hypoxia inducible factor-1α (Ad-HIF-1α-564/402/803).

METHODS:

Ad-HIF-1α-564/402/803 was amplified from HEK293A cells and purified by ultracentrifugation in CsCl gradient solutions. The infection efficiency was observed by X-gal staining. The lyophilized adenovirus was prepared under appropriate conditions. Before and after lyophilization, the effect of Ad-HIF-1α-564/402/803 on hMVEC proliferation was evaluated by MTS assay. The recombinant adenovirus was confirmed by PCR and DNA sequence analysis before and 1 day, 6 months and 12 months after lyophilization, and hMVECs infected with Ad-HIF-1α-564/402/803 at these time points were examined for HIF-1α protein expression using Western blotting.

RESULTS:

No significant changes were observed in the effect of lyophilized Ad-HIF-1α-564/402/803 on hMVECs proliferation at the optimal multiplicity of infection of 100 pfu/cell (P>0.05). At the 4 time points, the recombinant adenovirus HIF-1α showed no structural alterations or significant changes in the expression level of HIF-1α protein in the transfected hMVECs (P>0.05).

CONCLUSION:

Lyophilized Ad-HIF-1α-564/402/803 can maintain its biological activities for a long time.

• Authors: Yu TAO, Yue-gang WANG, Xuan WEI, Cheng LIU, Dong-dong CHEN, Ping-sheng WU
• Journal: Nan fang yi ke da xue xue bao = Journal of Southern Medical University (Nan Fang Yi Ke Da Xue Xue Bao) Vol. 31 Issue 3 Pg. 469-72 (Mar 2011) ISSN: 1673-4254 [Print] China
• PMID: 21421485 (Publication Type: English Abstract, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Monoclonal
• Hypoxia-Inducible Factor 1, alpha Subunit
• Mutant Proteins

Topics

• Adenoviridae (genetics, metabolism)
• Antibodies, Monoclonal (genetics)
• Freeze Drying
• Genetic Vectors
• HEK293 Cells
• Humans
• Hypoxia-Inducible Factor 1, alpha Subunit (biosynthesis, genetics)
• Mutant Proteins (genetics, metabolism)