Journal Article

Adenoviral targeting using genetically incorporated camelid single variable domains

Tremblay, Jacqueline M, Curiel, David T, Shoemaker, Charles B, Kaliberov, Sergey A, Kaliberova, Lyudmila N, Buggio, Maurizio

2014

Overview

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.

• Publisher: Nature Publishing Group US,Nature Publishing Group
• Subject: 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