Digoxigenin modification of adenovirus to spatially control gene delivery from chitosan surfaces
Digoxigenin modification of adenovirus to spatially control gene delivery from chitosan surfaces
复制标题
DOI：
10.1016/j.jconrel.2009.01.020
复制
发表时间：
2009-05-05
影响因子：
10.8
通讯作者：
Krebsbach, Paul H.
中科院分区：
文献类型：
--
作者：
Hu, Wei-Wen;Lang, Michael W.;Krebsbach, Paul H.
为了在空间上控制来自生物材料支架的多个病毒载体的递送，将地高辛（DIG）缀合至腺病毒衣壳蛋白作为用于抗体固定的抗原决定簇。通过对DIG修饰腺病毒的感染性、毒性、特异性和固定化稳定性的研究，探讨了DIG修饰腺病毒的可行性和有效性。在壳聚糖表面上缀合的抗DIG抗体能够与DIG修饰的腺病毒结合，并且可以稳定地结合在材料上至少两周，但是修饰足够温和，从而保持病毒感染性。为了鉴定两种不同的腺病毒，将蜡掩蔽应用于在壳聚糖膜的两个离散区域中缀合抗DIG和抗腺病毒抗体。在细胞培养后检查表达不同报告基因的这两种病毒载体的分布。来自转导细胞的荧光蛋白表达说明感染分布是可以控制的：一个基因被递送到生物材料的整个区域，另一个基因仅被递送到限定区域。与其他三种强心苷相比，当DIG缀合在腺病毒上时，ATP酶抑制不可检测，表明该方法对于体内应用可能是安全的。这种双重病毒载体递送系统应该能够在细胞信号传导病毒之间产生不同的界面，以控制来自一系列不同生物材料的组织再生。(c)2009爱思唯尔有限公司版权所有。
To spatially control the delivery of multiple viral vectors from biomaterial scaffolds, digoxigenin (DIG) was conjugated to adenoviral capsid proteins as an antigenic determinant for antibody immobilization. The infectivity, toxicity, specificity and immobilization stability of DIG-modified adenovirus were examined to investigate the feasibility and effectiveness of this viral surface modification. Anti-DIG antibody conjugated on chitosan surfaces was able to immobilize DIG-modified adenovirus and could be stably bound on the material for at least two weeks, yet the modification was mild enough that viral infectivity was maintained. To immobilize two different adenoviruses, wax masking was applied to conjugate anti-DIG and anti-adenovirus antibodies in two discrete regions of a chitosan film. The distribution of these two viral vectors expressing different reporter genes was examined after cell culture. Fluorescent protein expression from transduced cells illustrated that the infection distribution could be controlled: one gene was delivered to the entire region of the biomaterial, and another was only delivered to defined regions. Compared to three other cardiac glycosides, ATPase inhibition was undetectable when DIG was conjugated on the adenovirus, suggesting that the method may be safe for in vivo application. This dual viral vector delivery system should be capable of generating distinct interfaces between cell signaling viruses to control tissue regeneration from a range of different biomaterials. (c) 2009 Elsevier B.V. All rights reserved.