摘要
目的研究偶联单克隆抗体胶原涂覆的聚氨酯膜在基因定位递送中的应用。方法将抗腺病毒纤突的单克隆F(ab)’2抗体通过巯基反应共价键偶联到胶原涂覆的聚氨酯膜上,再通过抗体-抗原特异性反应携带病毒基因载体,得到局部定位的基因运载体系。用报告基因进行平滑肌细胞(A10细胞)的体外转染实验和模拟生理条件的稳定性实验。结果体外细胞转染实验显示,该方法携带的载基因病毒在A10细胞中实现了高效、高度定位的基因表达。模仿生理条件的体外稳定性实验表明,通过抗体结合在PU膜上的载基因病毒可缓慢持续地释放出来,有效地转染A10细胞达20d左右。结论这种经抗病毒抗体连接固定于胶原涂覆聚氨酯膜的病毒能很好地局部定位递送基因,可望用于临床基因治疗。
Objective To study the feasibility of delivering vital gene vector from a collagen-coated polyurethane （PU） film through a mechanism involving monoclonal antiviral antibody tethering. Methods Anti-adenoviral monoclonal antibodies were covalently bound to the collagen-coated PU surface. These antibodies enabled tethering of replication defective adenoviruses through highly specific antigen-antibody affinity. The PU film-based gene delivery using antibody-tethered adenovirus encoding green fluorescent protein （GFP） was tested in rat arterial smooth muscle cell （A10 cell） culture in vitro. The virus binding stability was studied by incubating the collagen-coated PU film in PBS solution at 37℃ for 20 days, followed with A10 cell cultures with the incubated films and the corresponding buffer solution. Results PU films with antibody-tethered adenovirus encoding GFP demonstrated efficient and highly localized gene delivery to A10 cells. Virus binding was stable for at least 10 days at physiological conditions, more than 77% of the originally bound virus remained in the film after 15 day＇s incubation. Conclusion Gene delivery using PU film-based anti-viral antibody tethering of vectors exhibited potentials of applications in a wide array of single or multiple therapeutic gene strategies, and in further stent-based gene delivery therapeutic strategies.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2006年第3期350-354,i0002,共6页
Acta Academiae Medicinae Sinicae
基金
国家自然科学基金(50473059)
天津科委重点基金(043803011)
高等学校博士点专项科研基金(20030023004)~~
