Systematic improvement of lentivirus transduction protocols by antibody fragments fused to VSV-G as envelope glycoprotein

Biomaterials. 2014 Apr;35(13):4204-12. doi: 10.1016/j.biomaterials.2014.01.051. Epub 2014 Feb 12.


Lentiviral vectors (LV) are widely used to successfully transduce cells for research and clinical applications. Lentiviral vectors pseudotyped with the vesicular stomatitis virus glycoprotein (VSV-G) can be produced to high titers and mediate high transduction efficiencies in vitro. For clinical applications the need for optimized transduction protocols and the limited activity of retronectin as LV enhancer, results in the application of a high multiplicity of infection (MOI) to achieve effective transduction efficiencies for a number of therapeutically relevant cells, e.g. CD34(+) hematopoietic stem cells, T- and B-cells. Our study describes an optimized LV infection protocol including a non-toxic poloxamer-based adjuvant combined with antibody-retargeted lentiviral particles, improving transduction efficiency at low MOI. Cell specificity of lentiviral vectors was increased by displaying different ratios of scFv-fused VSV-G glycoproteins on the viral envelope. The system was validated with difficult to transduce human CD30(+) lymphoma cells, and EGFR(+) tumor cells. Highly efficient transduction of lymphoma cells was achieved, >50% of cells were transduced when MOI 1 was used. The scFv displaying lentiviral particles gained relative specificity for transduction of target cells. Preferential gene delivery to CD30(+) or EGFR(+) cells was increased 4-fold in mixed cell cultures by presenting scFv antibody fragments binding to respective surface markers. A combination of spinoculation, poloxamer-based chemical adjuvant, and LV displaying scFv fragments increases transduction efficiencies of hard-to-transduce suspension lymphoma cells, and promises new chances for the future development of improved clinical protocols.

Keywords: Gene delivery; Gene transfer; Lentivirus; Poloxamer; VSV-G retargeting; scFv.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Gene Transfer Techniques
  • Genetic Vectors / genetics*
  • Glycoproteins / chemistry*
  • Humans
  • Immunoglobulin Fragments / chemistry*
  • Lentivirus / genetics*
  • Membrane Glycoproteins / chemistry*
  • Transduction, Genetic / methods*
  • Viral Envelope Proteins


  • Glycoproteins
  • Immunoglobulin Fragments
  • Membrane Glycoproteins
  • Viral Envelope Proteins