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Association of glycosylphosphatidylinositol‐anchored protein with retroviral particles
Author(s) -
Metzner Christoph,
Mostegl Meike M.,
Günzburg Walter H.,
Salmons Brian,
Dangerfield John A.
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.08-108217
Subject(s) - recombinant dna , group specific antigen , virus , flow cytometry , viral vector , cell culture , virology , biology , microbiology and biotechnology , chemistry , gene , biochemistry , genetics
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. FASEB J. 22, 2734–2739 (2008)