Comparison of adenovirus fiber, protein IX, and hexon capsomeres as scaffolds for vector purification and cell targeting
Comparison of adenovirus fiber, protein IX, and hexon capsomeres as scaffolds for vector purification and cell targeting
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DOI：
10.1016/j.virol.2006.01.032
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发表时间：
2006-06-05
期刊：
影响因子：
3.7
通讯作者：
Barry, Michael A.
中科院分区：
文献类型：
--
作者：
Campos, Samuel K.;Barry, Michael A.
用新的配体直接遗传修饰腺病毒衣壳蛋白是赋予腺病毒载体靶向性的有吸引力的手段。尽管已经报道了几种衣壳蛋白耐受外源肽和蛋白质的遗传融合，但是缺乏通过不同衣壳的细胞靶向效率的直接比较。同样地，由于缺乏可用于重靶向的captain相容性配体，尚未进行与一种或多种配体的直接比较。在这里，我们利用一组代谢生物素化的Ad载体直接比较靶向转导通过纤维，蛋白IX，和六邻体壳粒使用各种生物素化的配体，包括抗体，转铁蛋白，EGF，和霍乱毒素B。这些结果清楚地表明，用多种高亲和力受体结合配体靶向细胞仅在转导通过纤维蛋白重定向时有效。相比之下，蛋白IX和六邻体介导的靶向相同的一组配体未能介导强大的载体靶向，这可能是由于在细胞表面或靶向细胞内的异常运输。这些数据表明，通过基因掺入高亲和力配体的载体靶向可能通过修饰腺病毒纤维而不是蛋白IX和六邻体壳粒最有效。相比之下，使用代谢生物素化系统的Ad载体的单步单体抗生物素蛋白亲和纯化通过壳粒如蛋白IX和六邻体是最有效的。(c)2006年爱思唯尔公司All rights reserved.
The direct genetic modification of adenoviral capsid proteins with new ligands is an attractive means to confer targeted tropism to adenoviral vectors. Although several capsid proteins have been reported to tolerate the genetic fusion of foreign peptides and proteins, direct comparison of cell targeting efficiencies through the different capsomeres has been lacking. Likewise, direct comparison of with one or multiple ligands has not been performed due to a lack of capsid-compatible ligands available for retargeting. Here we utilize a panel of metabolically biotinylated Ad vectors to directly compare targeted transduction through the fiber, protein IX, and hexon capsomeres using a variety of biotinylated ligands including antibodies, transferrin, EGF, and cholera toxin B. These results clearly demonstrate that cell targeting with a variety of high affinity receptor-binding ligands is only effective when transduction is redirected through the fiber protein. In contrast, protein IX and hexon-mediated targeting by the same set of ligands failed to mediate robust vector targeting, perhaps due to aberrant trafficking at the cell surface or inside targeted cells. These data suggest that vector targeting by genetic incorporation of high affinity ligands will likely be most efficient through modification of the adenovirus fiber rather than the protein IX and hexon capsomeres. In contrast, single-step monomeric avidin affinity purification of Ad vectors using the metabolic biotinylation system is most effective through capsomeres like protein IX and hexon. (c) 2006 Elsevier Inc. All rights reserved.