The complexes of adenovirus and anionic liposomes: preparation and in vitro characterization.
- Author:
Zhi-Rong ZHONG
1
;
Yu WAN
;
San-Jun SHI
;
Zhi-Rong ZHANG
;
Xun SUN
Author Information
1. Western School of Pharmacy, Sichuan University, Chengdu 610041, China.
- Publication Type:Journal Article
- MeSH:
Adenoviridae;
genetics;
ultrastructure;
Animals;
Anions;
Cytopathogenic Effect, Viral;
Dogs;
Drug Compounding;
methods;
Genetic Vectors;
Green Fluorescent Proteins;
chemistry;
HEK293 Cells;
Humans;
Liposomes;
chemistry;
pharmacokinetics;
ultrastructure;
Madin Darby Canine Kidney Cells;
Microscopy, Confocal;
Microscopy, Electron, Transmission;
Particle Size;
Polymerase Chain Reaction;
methods;
Recombinant Fusion Proteins;
ultrastructure
- From:
Acta Pharmaceutica Sinica
2012;47(1):116-123
- CountryChina
- Language:Chinese
-
Abstract:
This study is to report the preparation of complexes of Ad5 and anionic liposomes (AL-Ad5), the amplification of adenoviruses with enhanced green fluorescent protein (eGFP) reporter gene performed by HEK 293 cells, the adenoviral vectors purified by cesium chloride gradient centrifugation, and the titer of adenovirus determined by cytopathic effect (CPE) method, hexon capsid immunoassay and quantitative-PCR (Q-PCR), separately. The prescription and experiment conditions were optimized by central composite design (CCD). The complexes of Ad5 and AL-Ad5 were formulated by the calcium-induced phase change method. The morpholopy, particle size and zeta potential were detected by dynamic light scattering (DLS) and transmission electron microscopy (TEM), respectively. Additionally, the bicolourable fluoresce-labeled complexes (F(labeled)-AL-Ad5) were prepared and their intracellular location in MDCK cells was detected by confocal laser scanning microscopy (CLSM). The results indicate that the complexes of AL-Ad5 exhibited a uniform distribution with a particle size of 211 +/- 10 nm and a zeta potential of -41.2 +/- 2.2 mV. The result of CLSM demonstrates that the intracellular location of red fluoresce-labeled adenovirus was consistent with that of green fluoresce-labeled liposomes suggesting that the naked adenovirus was well encapsulated by the anionic liposomes in complexes of AL-Ad5.
