The 'adenobody' approach to viral targeting: specific and enhanced adenoviral gene delivery.
Recombinant adenoviruses have enormous potential as vectors for gene therapy. They have evolved an efficient method of infection and a wide host range but this leads to concerns about the specificity of gene delivery. In order to target an adenovirus type 5-based vector we have investigated an antibody approach. A virus neutralising scFv antibody fragment was isolated from a phage library and a C-terminal fusion protein with epidermal growth factor (EGF) constructed. This fusion protein, or 'adenobody', bound both to the fibre protein of the adenovirus and to the EGF receptor (EGFR) on human cells, and was able to direct adenoviral binding to the new receptor. Using this system the efficiency of viral infection was markedly enhanced and was targeted to the EGFR. The adenobody-directed infection correlated with the level of EGF receptor expressed on the cells and could be blocked by competition with pure EGF. Peptide inhibition experiments suggest that infection is mediated directly through attachment to the EGFR and does not require penton-integrin interactions. This work shows that the 'adenobody' approach can enhance the efficiency as well as target adenoviral infection and has numerous potential applications for gene therapy.
Watkins
S J
SJ
Bristol University, Department of Oncology, Bristol Oncology Centre, UK.
Mesyanzhinov
V V
VV
Kurochkina
L P
LP
Hawkins
R E
RE
D000260:Adenoviruses, Human / Q000235:genetics* / Q000276:immunology; D000818:Animals; D000914:Antibodies, Viral / Q000378:metabolism*; D001435:Bacteriophages; D001667:Binding, Competitive; D004815:Epidermal Growth Factor / Q000378:metabolism*; D066246:ErbB Receptors / Q000378:metabolism; D018390:Gene Targeting / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D007128:Immunoglobulin Fragments / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000378:metabolism; D051379:Mice; D011993:Recombinant Fusion Proteins; D014407:Tumor Cells, Cultured / Q000378:metabolism
D000914:Antibodies, Viral; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D011993:Recombinant Fusion Proteins; D004815:Epidermal Growth Factor; D066246:ErbB Receptors