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Targeting of biotinylated compounds to its target tissue using a low-density lipoprotein receptor–avidin fusion protein

Gene Therapy volume 10, pages 2090–2097 (2003)Cite this article

Abstract

The very high binding affinity of avidin to biotin is one of the highest to occur in nature. We constructed a fusion protein composed of avidin and the endocytotic LDL receptor in order to target biotinylated molecules to cells of the desired tissues. In addition to the native avidin, charge-mutated and nonglycosylated avidins were utilized as part of the fusion proteins, in order to modify its properties. All of the fusion protein versions retained the biotin-binding capacity. Although the specificity was not increased, however, fusion proteins composed of natural avidin and nonglycosylated avidin bound most efficiently to the biotinylated ligands. Fluorescence microscopy and atomic force microscopy studies revealed the expression of the fusion protein on cell membranes, and demonstrated specific and high-affinity binding of biotin to the low-density lipoprotein receptor (LDLR)–avidin fusion protein in vitro. Additionally, systemically administered biotinylated ligand targeted with high specificity the intracerebral tumors of rats that were expressing fusion protein after the virus-mediated gene transfer. These results suggest that local gene transfer of the fusion protein to target tissues may offer a novel tool for the delivery of biotinylated molecules in vitro and in vivo for therapeutic and imaging purposes.

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Acknowledgements

This study was supported by the Finnish Academy, Kuopio University Hospital (EVO grant) and Ark Therapeutics Ltd. We thank Ms Piia Valonen MSc, Ms Irene Helkala, Ms Aila Erkinheimo, Ms Mervi Nieminen and Ms Seija Sahrio for the technical assistance, and Ms Marja Poikolainen for preparing the manuscript.

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  1. P Lehtolainen and T Wirth: Equal contribution.

Authors and Affiliations

  1. A.I. Virtanen Institute, University of Kuopio, Kuopio, Finland

    P Lehtolainen, T Wirth, A K Taskinen, O Leppänen, M Lappalainen, K Pulkkanen, K J Airenne & S Ylä-Herttuala

  2. Department of Medicine, Bone and Mineral Centre, The Rayne Institute, London, UK

    P Lehenkari & M Horton

  3. University of Jyväskylä, Jyväskylä, Finland

    A Marttila, V Marjomäki & M S Kulomaa

  4. Department of Medicine, University of Kuopio and Gene Therapy Unit, Kuopio, University Hospital, Kuopio, Finland

    S Ylä-Herttuala

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  1. P Lehtolainen
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Lehtolainen, P., Wirth, T., Taskinen, A. et al. Targeting of biotinylated compounds to its target tissue using a low-density lipoprotein receptor–avidin fusion protein. Gene Ther 10, 2090–2097 (2003). https://doi.org/10.1038/sj.gt.3302120

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