Selection of muscle-binding peptides from context-specific peptide-presenting phage libraries for adenoviral vector targeting
Selection of muscle-binding peptides from context-specific peptide-presenting phage libraries for adenoviral vector targeting
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DOI：
10.1128/jvi.79.21.13667-13672.2005
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发表时间：
2005-11-01
影响因子：
5.4
通讯作者：
Barry, MA
中科院分区：
文献类型：
--
作者：
Ghosh, D;Barry, MA
细胞靶向载体的生产部分涉及向载体添加新的靶向配体，以介导与目标细胞的结合。对于病毒载体，理想的方法是将新的配体基因工程改造到病毒的衣壳蛋白中，以产生一种单一的介导治疗的制剂。尽管这是理想的，但将一种外源性配体从一种结构环境插入到衣壳蛋白的不同结构环境中可能会消除配体的功能或破坏病毒的组装和功能。为了解决腺病毒载体的这种环境问题，我们构建了一个“环境特异性”的呈肽噬菌体文库。我们在fd噬菌体的pIII蛋白上展示了一个12个氨基酸（12聚体）的随机肽文库，该文库位于5型腺病毒纤维蛋白的H和I片层之间。这个文库被用于针对C2C12小鼠骨骼肌细胞的肽筛选。经过五轮筛选以及在非目标细胞上进行四轮清除，筛选出一种主要的肽，命名为12.51，它与目标C2C12细胞的结合能力比阳性对照RGD肽大约强100倍。将12.51翻译回纤维蛋白产生了一种配体修饰的腺病毒载体，它介导对目标C2C12细胞的转导能力提高了14倍。这些数据表明，环境特异性的呈肽文库可能允许选择相容的肽配体，以便功能性地翻译到病毒载体中用于重新靶向。
Production of cell-targeting vectors in part involves the addition of new targeting ligands to the vector to mediate binding to the cells of interest. For viral vectors, the ideal approach is to genetically engineer new ligands into the capsid proteins of the virus to generate a single agent to mediate therapy. Although this is ideal, this insertion of an exogenous ligand from one structural context into the differing structural context of a capsid protein can ablate the function of the ligand or disrupt viral assembly and function. To address this context problem for adenoviral vectors, we have engineered a "context-specific" peptide-presenting phage library. We have displayed a 12-amino-acid (12-mer) random peptide library between the H and I sheets of the fiber protein of adenovirus type 5 on the pIII protein of fd bacteriophage. This library was used for peptide selection against C2C12 mouse skeletal muscle cells. Five rounds of selection combined with four rounds of clearing on nontarget cells selected one primary peptide designated 12.51, which bound target C2C12 cells approximately 100-fold better than the positive control RGD peptide. Translation of 12.51 back into the fiber protein produced a ligand-modified adenoviral vector that mediated 14-fold-better transduction of target C2C12 cells. These data suggest context-specific peptide-presenting libraries may allow selection of compatible peptide ligands for functional translation into viral vectors for retargeting.