Targeting lentiviral vector to specific cell types through surface displayed single chain antibody and fusogenic molecule.
BACKGROUND
Viral 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.


RESULTS
Two 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 (alphaCD20) 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.


CONCLUSIONS
Certain 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.
Lei
Yuning
Y
Mork Family Department of Chemical Engineering and Materials Science, University of Southern California, Los Angeles, CA 90089, USA.
Joo
Kye-Il
KI
Zarzar
Jonathan
J
Wong
Clement
C
Wang
Pin
P
D002460:Cell Line; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics / Q000502:physiology*; D057127:Single-Chain Antibodies / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic; D014760:Viral Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D056189:Viral Tropism; D053585:Virus Attachment; D053586:Virus Internalization
D057127:Single-Chain Antibodies; D014760:Viral Fusion Proteins