Abstract
The unique ability of human adenovirus serotype 5 (Ad5) to accomplish efficient transduction has allowed the use of Ad5-based vectors for a range of gene therapy applications. Several strategies have been developed to alter tropism of Ad vectors to achieve a cell-specific gene delivery by using fiber modifications via genetic incorporation of targeting motifs. In this study, we have explored the utility of novel anti-human carcinoembryonic antigen (hCEA) single variable domains derived from heavy chain (VHH) camelid family of antibodies to achieve targeted gene transfer. To obtain anti-CEA VHHs, we produced a VHH-display library from peripheral blood lymphocytes RNA of alpacas at the peak of immune response to the hCEA antigen (Ag). We genetically incorporated an anti-hCEA VHH into a de-knobbed Ad5 fiber-fibritin chimera and demonstrated selective targeting to the cognate epitope expressed on the membrane surface of target cells. We report that the anti-hCEA VHH used in this study retains Ag recognition functionality and provides specificity for gene transfer of capsid-modified Ad5 vectors. These studies clearly demonstrated the feasibility of retargeting of Ad5-based gene transfer using VHHs.
Publication types
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Comparative Study
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Research Support, N.I.H., Extramural
MeSH terms
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Adenoviridae / physiology*
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Animals
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Antibody Specificity
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Camelids, New World*
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Capsid Proteins / administration & dosage
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Capsid Proteins / genetics
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Capsid Proteins / metabolism
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Carcinoembryonic Antigen / chemistry
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Carcinoembryonic Antigen / metabolism*
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Cell Line
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Cell Line, Tumor
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Feasibility Studies
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Gene Transfer Techniques*
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Genetic Vectors / administration & dosage
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Genetic Vectors / physiology*
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Humans
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Immunoglobulin Heavy Chains / administration & dosage
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Immunoglobulin Heavy Chains / chemistry
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Immunoglobulin Heavy Chains / genetics
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Immunoglobulin Heavy Chains / metabolism
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Immunoglobulin Variable Region / administration & dosage*
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Immunoglobulin Variable Region / chemistry
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Immunoglobulin Variable Region / genetics
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Immunoglobulin Variable Region / metabolism
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Male
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Peptide Fragments / administration & dosage
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Peptide Fragments / chemistry
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Peptide Fragments / genetics
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Peptide Fragments / metabolism
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Recombinant Fusion Proteins / administration & dosage
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / metabolism
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Recombinant Proteins / administration & dosage
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Recombinant Proteins / chemistry
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Recombinant Proteins / metabolism
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Transduction, Genetic
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Viral Proteins / administration & dosage
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Viral Proteins / genetics
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Viral Proteins / metabolism
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Viral Tropism*
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Virion / physiology
Substances
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Capsid Proteins
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Carcinoembryonic Antigen
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Immunoglobulin Heavy Chains
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Immunoglobulin Variable Region
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Peptide Fragments
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Recombinant Fusion Proteins
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Recombinant Proteins
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Viral Proteins
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fibritin protein, Enterobacteria phage T4