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This Article Full Text Full Text (PDF) All Versions of this Article: fj.08-108217v1 22/8/2734    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Google Scholar Articles by Metzner, C. Articles by Dangerfield, J. A. PubMed PubMed Citation Articles by Metzner, C. Articles by Dangerfield, J. A.

Association of glycosylphosphatidylinositol-anchored protein with retroviral particles

Christoph Metzner^*,,1, Meike M. Mostegl^*,, Walter H. Günzburg^*,,, Brian Salmons^,|| and John A. Dangerfield^*,,||

^* Institute of Virology, University of Veterinary Medicine, Vienna, Austria;

Christian Doppler Laboratory for Gene Therapeutic Vector Development, Vienna, Austria;

Christian Doppler Laboratory for Gene Therapeutic Vector Development, Foreign Module for Retroviral Nanotechnology, Singapore;

Austrianova Biomanufacturing, Vienna, Austria; and

^|| Austrianova Singapore, Singapore

¹Correspondence: Institute of Virology, University of Veterinary Medicine, Veterinärplatz 1, A-1210 Vienna, Austria. E-mail: christoph.metzner{at}vu-wien.ac.at

We describe for the first time the association of glycosylphosphatidylinositol (GPI) -anchored proteins with retroviral and lentiviral particles, similar to a process well established for cells, termed "painting." The aim of the study was to assess the feasibility of modification of retroviral vectors by exogenous addition of recombinant protein, removing the need for genetic engineering of virus producer cell lines. The recombinant GPI protein CD59his was purified via fast protein liquid chromatography and associated with concentrated virus stock in a controlled incubation procedure. Reaction mixtures were purified in order to remove nonassociated GPI protein and endogenous protein. Analysis of samples by immunoblotting revealed that CD59his was only detectable in the presence of viral particles. From this, we conclude that CD59his could be stably associated with retroviral particles. In addition, we demonstrated by flow cytometry that virus particles remain infectious after these procedures. As well as suggesting a novel possibility for interaction between enveloped virus and host, we believe that the stable association of recombinant GPI proteins to retroviral particles can be developed into an important tool for both research and clinical applications, especially in the fields of gene therapy and vaccine development.—Metzner, C., Mostegl, M. M., Günzburg, W. H., Salmons, B., Dangerfield, J. A. Association of glycosylphosphatidylinositol-anchored protein with retroviral particles.

Key Words: GPI • retrovirus • lentivirus • gene therapy • vaccine development