Dobroff Andrey S, Rangel Roberto, Guzman-Roja Liliana, Salmeron Carolina C, Gelovani Juri G, Sidman Richard L, Bologa Cristian G, Oprea Tudor I, Brinker C Jeffrey, Pasqualini Renata, Arap Wadih
Division of Hematology/Oncology, Department of Internal Medicine, University of New Mexico School of Medicine, Albuquerque, New Mexico.
Division of Molecular Medicine, Department of Internal Medicine, University of New Mexico School of Medicine, Albuquerque, New Mexico.
Curr Protoc Protein Sci. 2015 Feb 2;79:30.4.1-30.4.30. doi: 10.1002/0471140864.ps3004s79.
Phage display is a resourceful tool to, in an unbiased manner, discover and characterize functional protein-protein interactions, create vaccines, and engineer peptides, antibodies, and other proteins as targeted diagnostic and/or therapeutic agents. Recently, our group has developed a new class of internalizing phage (iPhage) for ligand-directed targeting of organelles and to identify molecular pathways within live cells. This unique technology is suitable for applications ranging from fundamental cell biology to drug development. This unit describes the methods for generating and screening the iPhage display system, and explains how to select and validate candidate internalizing homing peptide.
噬菌体展示是一种有效的工具,可用于以无偏见的方式发现和表征功能性蛋白质-蛋白质相互作用、开发疫苗以及设计肽、抗体和其他蛋白质作为靶向诊断和/或治疗剂。最近,我们团队开发了一类新型内化噬菌体(iPhage),用于细胞器的配体导向靶向,并识别活细胞内的分子途径。这项独特的技术适用于从基础细胞生物学到药物开发的广泛应用。本单元介绍了生成和筛选iPhage展示系统的方法,并解释了如何选择和验证候选内化归巢肽。