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Title: Comparison of adenovirus fiber, protein IX, and hexon capsomeres as scaffolds for vector purification and cell targeting

Abstract

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 andmore » 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.« less

Authors:
 [1];  [2]
  1. Department of Biochemistry and Cell Biology, Rice University, Houston, TX 77005 (United States)
  2. Department of Bioengineering, Rice University, Houston, TX 77005 (United States) and Center for Cell and Gene Therapy, Baylor College of Medicine, Methodist Hospital, and Texas Children's Hospital, Houston, TX 77030 (United States) and Departments of Immunology and Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030 (United States)
Publication Date:
OSTI Identifier:
20850518
Resource Type:
Journal Article
Journal Name:
Virology
Additional Journal Information:
Journal Volume: 349; Journal Issue: 2; Other Information: DOI: 10.1016/j.virol.2006.01.032; PII: S0042-6822(06)00042-0; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0042-6822
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ADENOVIRUS; AFFINITY; ANTIBODIES; AVIDIN; CHOLERA; DISEASE VECTORS; FIBERS; LIGANDS; PEPTIDES; PURIFICATION; RECEPTORS; TOXINS; TRANSFERRIN

Citation Formats

Campos, Samuel K, Center for Cell and Gene Therapy, Baylor College of Medicine, Methodist Hospital, and Texas Children's Hospital, Houston, TX 77030, and Barry, Michael A. Comparison of adenovirus fiber, protein IX, and hexon capsomeres as scaffolds for vector purification and cell targeting. United States: N. p., 2006. Web. doi:10.1016/j.virol.2006.01.032.
Campos, Samuel K, Center for Cell and Gene Therapy, Baylor College of Medicine, Methodist Hospital, and Texas Children's Hospital, Houston, TX 77030, & Barry, Michael A. Comparison of adenovirus fiber, protein IX, and hexon capsomeres as scaffolds for vector purification and cell targeting. United States. https://doi.org/10.1016/j.virol.2006.01.032
Campos, Samuel K, Center for Cell and Gene Therapy, Baylor College of Medicine, Methodist Hospital, and Texas Children's Hospital, Houston, TX 77030, and Barry, Michael A. 2006. "Comparison of adenovirus fiber, protein IX, and hexon capsomeres as scaffolds for vector purification and cell targeting". United States. https://doi.org/10.1016/j.virol.2006.01.032.
@article{osti_20850518,
title = {Comparison of adenovirus fiber, protein IX, and hexon capsomeres as scaffolds for vector purification and cell targeting},
author = {Campos, Samuel K and Center for Cell and Gene Therapy, Baylor College of Medicine, Methodist Hospital, and Texas Children's Hospital, Houston, TX 77030 and Barry, Michael A},
abstractNote = {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.},
doi = {10.1016/j.virol.2006.01.032},
url = {https://www.osti.gov/biblio/20850518}, journal = {Virology},
issn = {0042-6822},
number = 2,
volume = 349,
place = {United States},
year = {Mon Jun 05 00:00:00 EDT 2006},
month = {Mon Jun 05 00:00:00 EDT 2006}
}