Fabrication of surface functionalized adenovirus nano-carriers for intravascular injection

Abstract

[한글]Adenovirus (Ad) vectors are promising candidates for both gene transfer and vaccine applications. Chitosan is presently under investigation for many biomedical applications. Electrospinning can easily produce nano-structured synthetic polymer. Folic acid (FA) improved gene transfection efficiency by promoting the uptake of folate receptor bearing cancer cells.Surface functionalized Ad nano-carriers with low-molecular chitosan (LMC), polyethyleneglycol (PEG), and FA were fabricated successfully by electrospinning method for intravascular injection. At first, pristine Ad was encapsulated with LMC by using electrospinning method. The size of the LMC coated Ad (ADV-cLMC) is about 70 nm that would be enough to be used for intravascular injection. To confirm the LMC coating of pristine Ad, surface charges of pristine Ad and ADV-cLMC were measured by ζ -potential analysis. The ζ-potential of Ad and ADV-cLMC were -5.0 ± 2.0 mV and 3.3 ± 2.0 mV, respectively. Morphology of the ADV-cLMC was observed by transmission electron microscope (TEM). In the second step, PEG-FA synthesized in advance was conjugated with ADV-cLMC. The PEG-FA conjugated ADV-cLMC (ADV-cLMC-PEG-FA, ADV-LCPF) exhibited long circulation time and high solubility due to PEG conjugation. The surface functionalized Ad nano-carriers also had cancer targeting moiety due to FA modification. The ADV-LCPF was characterized by using fast protein liquid chromatography and gel electrophoresis. The size of final ADV- LCPF nano-carriers was about 150 ~ 200 nm. Cell viability was according to various FA concentration GFP expression measured by FACS

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