Efficient incorporation of a functional hyper-stable single-chain antibody fragment protein-IX fusion in the adenovirus capsid
| Autor: | J de Vrij, Jort Vellinga, Taco Gilles Uil, Pierre Martineau, Leif Lindholm, Rob C. Hoeben, S. Myhre |
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| Přispěvatelé: | Le Ster, Yves, Department of Molecular Cell Biology, Leiden University Medical Center (LUMC), Got-a-Gene AB, Got-A-Gene AB, Institut de Recherche en Infectiologie de Montpellier (IRIM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), We thank Ronald W.A.L. Limpens (Leiden University Medical Center) for help with immuno-affinity electron microscopy, and participants in the GIANT program for stimulating discussions. This work was supported by the Technology Foundation STW (program LGN66.3977), and the European Union through the 6th Framework Program GIANT (Contract no.: 512087). |
| Jazyk: | angličtina |
| Rok vydání: | 2007 |
| Předmět: |
Genetic enhancement
Recombinant Fusion Proteins viruses Blotting Western Genetic Vectors Gene Expression Gene delivery medicine.disease_cause Article Viral vector Adenoviridae Fusion gene 03 medical and health sciences 0302 clinical medicine Transduction Genetic Genetics medicine capsid [SDV.BBM] Life Sciences [q-bio]/Biochemistry Molecular Biology Humans [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Microscopy Immunoelectron Molecular Biology Immunoglobulin Fragments targeting 030304 developmental biology Cell Line Transformed [SDV.IB] Life Sciences [q-bio]/Bioengineering 0303 health sciences biology Lentivirus Genetic Therapy adenovirus protein IX beta-Galactosidase Fusion protein Molecular biology Immunohistochemistry Capsid 030220 oncology & carcinogenesis hyper-stable scFv biology.protein Molecular Medicine Capsid Proteins [SDV.IB]Life Sciences [q-bio]/Bioengineering Antibody Genetic Engineering |
| Zdroj: | Gene Therapy Gene Therapy, 2007, 14 (8), pp.664-70. ⟨10.1038/sj.gt.3302908⟩ Gene Therapy, Nature Publishing Group, 2007, 14 (8), pp.664-70. ⟨10.1038/sj.gt.3302908⟩ |
| ISSN: | 0969-7128 1476-5462 |
| DOI: | 10.1038/sj.gt.3302908⟩ |
| Popis: | International audience; Recombinant adenoviruses are frequently used as gene transfer vehicles for therapeutic gene delivery. Strategies to amend their tropism include the incorporation of polypeptides with high affinity for cellular receptors. Single-chain antibodies have a great potential to achieve such cell type specificity. In this study, we evaluated the efficiency of incorporation of a single-chain antibody fused with the adenovirus minor capsid protein IX in the capsid of adenovirus type 5 vectors. To this end, the codons for the single-chain antibody fragments (scFv) 13R4 were fused with those encoding of pIX via a 75-Angstrom spacer sequence. The 13R4 is a hyper-stable single-chain antibody directed against beta-galactosidase, which was selected for its capacity to fold correctly in a reducing environment such as the cytoplasm. A lentiviral vector was used to stably express the pIX.flag.75.13R4.MYC.HIS fusion gene in 911 helper cells. Upon propagation of pIX-gene deleted human adenovirus-5 vectors on these cells, the pIX-fusion protein was efficiently incorporated in the capsid. Here, the 13R4 scFv was functional as was evident from its capacity to bind its ligand beta-galactosidase. These data demonstrate that the minor capsid protein IX can be used as an anchor for incorporation of single-chain antibodies in the capsids of adenovirus vectors. |
| Databáze: | OpenAIRE |
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