title: [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).
  
  
  CONCLUSIONS
  Lyophilized Ad-HIF-1α-564/402/803 can maintain its biological activities for a long time.
authors.affiliation: Department of Cardiology, Nanfang hospital, Southern medical University Guangzhou, 510515, China.E-mail: taoyu93@163.com.
authors.forename: Yu
authors.initials: Y
authors.lastname: TAO
authors.forename: Yue-gang
authors.initials: YG
authors.lastname: WANG
authors.forename: Xuan
authors.initials: X
authors.lastname: WEI
authors.forename: Cheng
authors.initials: C
authors.lastname: LIU
authors.forename: Dong-dong
authors.initials: DD
authors.lastname: CHEN
authors.forename: Ping-sheng
authors.initials: PS
authors.lastname: WU
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000235:genetics; D005612:Freeze Drying; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000096:biosynthesis* / Q000235:genetics; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D050505:Mutant Proteins