Abstract
BackgroundViral delivery remains one of the most commonly used techniques today in the field of gene therapy. However, one of the remaining hurdles is the off-targeting effect of viral delivery. To overcome this obstacle, we recently developed a method to incorporate an antibody and a fusogenic molecule (FM) as two distinct molecules into the lentiviral surface. In this report, we expand this strategy to utilize a single chain antibody (SCAb) for targeted transduction.ResultsTwo versions of the SCAb were generated to pair with our various engineered FMs by linking the heavy chain and the light chain variable domains of the anti-CD20 antibody (αCD20) via a GS linker and fusing them to the hinge-CH2-CH3 region of human IgG. The resulting protein was fused to either a HLA-A2 transmembrane domain or a VSVG transmembrane domain for anchoring purpose. Lentiviral vectors generated with either version of the SCAb and a selected FM were then characterized for binding and fusion activities in CD20-expressing cells.ConclusionCertain combinations of the SCAb with various FMs could result in an increase in viral transduction. This two-molecule lentiviral vector system design allows for parallel optimization of the SCAb and FMs to improve targeted gene delivery.
Highlights
Gene therapy is the introduction of a functional gene into a dysfunctional cell for a therapeutic benefit
In order to expand the targeted specificity of viral vectors beyond their natural tropism, numerous studies have been focused on pseudotyping lentiviral vectors with envelope glycoproteins derived from other viruses, such as the glycoprotein from vesicular stomatitis virus (VSVG) [3,4]
We report our attempt to utilize a single chain antibody (SCAb) to pair with a fusogenic molecule (FM) for targeting lentiviral vectors
Summary
Gene therapy is the introduction of a functional gene into a dysfunctional cell for a therapeutic benefit. Since the VSVG is thought to recognize a ubiquitous membrane phospholipids instead of a unique cellular receptor, pseudotyping generates vectors with broad specificities [5,6] To mitigate this off-target effect, previous attempts have been devoted to engineer the viral glycoprotein to recognize a specific cellular target by insertion of ligands, peptides, or antibodies [7,8,9,10,11,12,13,14,15,16]. We expand this strategy to utilize a single chain antibody (SCAb) for targeted transduction
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