Targeting of vaccinia virus using biotin-avidin viral coating and biotinylated antibodies

J Surg Res. 2005 Jan;123(1):49-54. doi: 10.1016/j.jss.2004.04.022.


Introduction: To test a general method for altering the tropism of viral vectors, we conjugated targeting antibody to the surface of recombinant vaccinia virus with a biotin-avidin-biotin linker and assessed the resulting infectivity in target cells and controls.

Materials and methods: We biotinylated a vaccinia viral vector and used avidin to crosslink the biotinylated viral surface to a biotinylated antibody specific for a molecule on the surface of a target cell. In an in vitro model system, we coated a recombinant vaccinia construct containing the E. coli beta-galactosidase gene with antibody to the murine class I MHC molecule Db. Target cells were B78H1 murine melanoma cells transduced with either the Db gene or, as a control, the Kb gene. Infectivity was assessed by staining target cells with x-gal to demonstrate expression of virally delivered beta-galactosidase. This technique was also assessed in a second system with vaccinia/beta-gal targeted to the murine B7.2 molecule. The infectivity of the resulting construct was assessed for murine SA1 fibrosarcoma cells transfected with the B7.2 gene and for wild-type, B7.2-negative SA1. Experiments were repeated in each system with similar results.

Results: This strategy demonstrated antibody-mediated viral targeting in both the B78H1 and the SA1 models. Importantly, addition of the targeting coat diminished the infectivity of the modified vaccinia for control cells but preserved infectivity for targeted cells. In the B78H1 system, Db-targeted vaccinia consistently had 2- to 3-fold greater infectivity for B78H1Db than B78H1Kb. Increasing the number of avidin molecules used per virion in the synthesis of the viral coat led to greater selectivity but decreased overall infectivity. In the SA1 system, B7.2-targeted vaccinia demonstrated completely ablated infectivity for control SA1 cells, but maintained infectivity for target SA1/B7.2 cells.

Conclusions: Recombinant viral vectors such as vaccinia may be coated with biotin/avidin and linked to biotinylated antibodies to preferentially target specific cell types in vitro. Such an approach may be useful in targeting recombinant lytic viruses to tumors for destruction and in immune up-regulation in vivo. Similarly, this approach may enhance nonlytic viruses for gene therapy applications.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / metabolism*
  • Antigens, CD / metabolism
  • Avidin / metabolism*
  • B7-2 Antigen
  • Biotin / metabolism*
  • Biotinylation
  • Genetic Therapy / methods*
  • Genetic Vectors
  • Histocompatibility Antigens Class I / physiology
  • Membrane Glycoproteins / metabolism
  • Mice
  • Tropism
  • Vaccinia virus / genetics*
  • Vaccinia virus / immunology
  • Vaccinia virus / physiology


  • Antibodies, Monoclonal
  • Antigens, CD
  • B7-2 Antigen
  • Cd86 protein, mouse
  • Histocompatibility Antigens Class I
  • Membrane Glycoproteins
  • Avidin
  • Biotin