Adenoviral targeting using genetically incorporated camelid single variable domains

• Published in: Laboratory investigation 2014-08, Vol.94 (8), p.893-905
• Main Authors: Kaliberov, Sergey A, Kaliberova, Lyudmila N, Buggio, Maurizio, Tremblay, Jacqueline M, Shoemaker, Charles B, Curiel, David T
• Format: Article
• Language: English
• Subjects: 631/1647/2300/1514; 631/61/201; Adenoviridae - physiology; Animals; Antibody Specificity; Camelids, New World; Capsid Proteins - administration & dosage; Capsid Proteins - genetics; Capsid Proteins - metabolism; Carcinoembryonic Antigen - chemistry; Carcinoembryonic Antigen - metabolism; Cell Line; Cell Line, Tumor; Feasibility Studies; Gene Transfer Techniques; Genetic Vectors - administration & dosage; Genetic Vectors - physiology; Human adenovirus; Humans; Immunoglobulin Heavy Chains - administration & dosage; Immunoglobulin Heavy Chains - chemistry; Immunoglobulin Heavy Chains - genetics; Immunoglobulin Heavy Chains - metabolism; Immunoglobulin Variable Region - administration & dosage; Immunoglobulin Variable Region - chemistry; Immunoglobulin Variable Region - genetics; Immunoglobulin Variable Region - metabolism; Laboratory Medicine; Male; Medicine; Medicine & Public Health; Pathology; Peptide Fragments - administration & dosage; Peptide Fragments - chemistry; Peptide Fragments - genetics; Peptide Fragments - metabolism; Recombinant Fusion Proteins - administration & dosage; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - metabolism; Recombinant Proteins - administration & dosage; Recombinant Proteins - chemistry; Recombinant Proteins - metabolism; research-article; Transduction, Genetic; Viral Proteins - administration & dosage; Viral Proteins - genetics; Viral Proteins - metabolism; Viral Tropism; Virion - physiology
• ISSN: 0023-6837; 1530-0307
• DOI: 10.1038/labinvest.2014.82

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.