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Antibody Modified Collagen Matrix for Site-Specific Gene Delivery

文献类型: 会议论文

第一作者: Xu Jin

作者: Xu Jin 1 ; Lanxia Liu 1 ; Lin Mei 1 ; Xigang Leng 1 ; Chao Zhang 1 ; Hongfan Sun 1 ; Cunxian Song 1 ; R.J. Levy 2 ;

作者机构: 1.Institute of Biomedical Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, The Tianjin Key Laboratory of Biomaterial Research, Tianjin 300192, People's Republic of China

2.The Children's Hospital of Philadelphia, Abramson Pediatric Research Center, Suite 702, 3516 Civic Center Boulevard, Philadelphia, PA 19104

关键词: Antibody;Collagen Matrix;Specific Gene Delivery

会议名称: Pacifichem 2005

主办单位:

页码: 243-264

摘要: We investigated the feasibility of immobilizing plasmid DNA on collagen matrix through covalently coupled anti-DNA antibody to achieve long-lasting and site-specific DNA delivery. Methods: Collagen was modified with SPDP (Nsuccinimidyl-3- (2-pyridyldithiol)-propionate) / DTT to introduce –SH group on collagen matrix. A pyridyl disulfide modified anti-DNA IgM wasthen covalently linked with collagen through thiol reaction. These antibodies enabled tethering of plasmid DNA through highly specific antigen-antibody affinity. Radioactive-labelled antibody and pDNA were used to evaluate binding capacity and stability. A plasmid EGFP-N3 was tethered on anti-DNA antibody-modified collagen-coated coronary stent as model gene delivery system that was assessed in both cell culture and in a rabbit model. Results showed that the amount of antibody bound on collagen films was 15 times higher than that of the control films (without SPDP chemical linking), and that the retention time of the antibody on films was also significantly longer. The pDNA-tethered and collagen-coated stents had no detrimental effect on cell growth. In cell culture studies, numerous GFP-transduced cells were detected only on the stent surface, thus demonstrating efficient and highly localized gene delivery. The overall GFP transduction efficiency in treated rabbit coronary arteries was 2.8±0.7% of the total cells. However, the rate of neointima transduction was about 7% of total cells in this region. Importantly, no distal spread of the vector was detectable by PCR, either in distal organs, or in the downstream segments of the stented arteries. In conclusion, we report the first successful use of anti-DNA antibody-modified stent as plasmid gene delivery system; this technique thus represents an efficient and highly localized gene delivery system both in vitro and in vivo.

分类号: R965

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