Ligand-directed profiling of organelles with internalizing phage libraries

Andrey S Dobroff; Roberto Rangel; Liliana Guzman-Roja; Carolina C Salmeron; Juri G Gelovani; Richard L Sidman; Cristian G Bologa; Tudor I Oprea; C Jeffrey Brinker; Renata Pasqualini; Wadih Arap

doi:10.1002/0471140864.ps3004s79

Ligand-directed profiling of organelles with internalizing phage libraries

Curr Protoc Protein Sci. 2015 Feb 2:79:30.4.1-30.4.30. doi: 10.1002/0471140864.ps3004s79.

Authors

Andrey S Dobroff^{1

2

3}, Roberto Rangel^{4

3}, Liliana Guzman-Roja^{4

3}, Carolina C Salmeron^{1

2}, Juri G Gelovani⁵, Richard L Sidman⁶, Cristian G Bologa⁷, Tudor I Oprea⁷, C Jeffrey Brinker⁸, Renata Pasqualini^{2

9}, Wadih Arap^{1

9}

Affiliations

¹ Division of Hematology/Oncology, Department of Internal Medicine, University of New Mexico School of Medicine, Albuquerque, New Mexico.
² Division of Molecular Medicine, Department of Internal Medicine, University of New Mexico School of Medicine, Albuquerque, New Mexico.
³ These authors contributed equally to this work.
⁴ Cancer Research Program, Houston Methodist Research Institute, Houston, Texas.
⁵ Department of Biomedical Engineering, Wayne State University, Detroit, Michigan.
⁶ Department of Neurology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts.
⁷ Translational Informatics Division, Department of Internal Medicine, University of New Mexico School of Medicine, Albuquerque, New Mexico.
⁸ Department of Chemical and Nuclear Engineering, The University of New Mexico Cancer Center, Albuquerque, New Mexico.
⁹ These authors contributed equally as senior authors to this work.

Abstract

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.

Keywords: intracellular receptors; intracellular targeting; mammalian cells; organelles; penetratin; peptides; phage display; proteomics.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Cell Line, Tumor
Cell Surface Display Techniques / methods*
Cloning, Molecular
Cytological Techniques / methods*
Escherichia coli
Humans
Organelles / chemistry*
Organelles / metabolism*

Grants and funding

U54 CA151668/CA/NCI NIH HHS/United States