Metabolically biotinylated adenovirus for cell targeting, ligand screening, and vector purification

M. Brandon Parrott, Kristen E. Adams, George T. Mercier, Hoyin Mok, Samuel K Campos, Michael A. Barry

Research output: Contribution to journalArticle

82 Citations (Scopus)

Abstract

Development of cell-targeting vectors is an important focus for gene therapy. While some ligands can be genetically inserted into virus capsid proteins for cell targeting, for many ligands, this approach can disrupt either ligand function or vector function. To address this problem for adenovirus type 5 vectors, the fiber capsid protein was genetically fused to a biotin acceptor peptide (BAP). Adenovirus particles bearing this BAP were metabolically biotinylated during vector production by the endogenous biotin ligase in 293 cells to produce covalently biotinylated virions. The resulting biotinylated vector could be retargeted to new receptors by conjugation to biotinylated antibodies using tetrameric avidin (Kd = 10-15 M). The biotinylated vector could also be purified by biotin-reversible binding on monomeric avidin (Kd = 10-7 M). Finally, this vector was used as a ligand screening platform for dendritic cells in which a variety of structurally diverse protein, carbohydrate, and nucleic acid ligands were easily added to the vector using the biotin-avidin interaction. This work demonstrates the utility of metabolically biotinylated viruses for ligand screening, vector targeting, and virus purification applications.

Original languageEnglish (US)
Pages (from-to)688-700
Number of pages13
JournalMolecular Therapy
Volume8
Issue number4
DOIs
StatePublished - Oct 2003
Externally publishedYes

Fingerprint

Adenoviridae
Biotin
Ligands
Avidin
Capsid Proteins
Viruses
Peptides
Protein Transport
Ligases
Genetic Therapy
Virion
Dendritic Cells
Nucleic Acids
Carbohydrates
Antibodies
Proteins

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Metabolically biotinylated adenovirus for cell targeting, ligand screening, and vector purification. / Parrott, M. Brandon; Adams, Kristen E.; Mercier, George T.; Mok, Hoyin; Campos, Samuel K; Barry, Michael A.

In: Molecular Therapy, Vol. 8, No. 4, 10.2003, p. 688-700.

Research output: Contribution to journalArticle

Parrott, M. Brandon ; Adams, Kristen E. ; Mercier, George T. ; Mok, Hoyin ; Campos, Samuel K ; Barry, Michael A. / Metabolically biotinylated adenovirus for cell targeting, ligand screening, and vector purification. In: Molecular Therapy. 2003 ; Vol. 8, No. 4. pp. 688-700.
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