Abstract
The construction protocol of bio-nanocapsule (BNC)-based nanocarriers, named GL-BNC and GL-virosome, for targeted drug delivery to macrophages is described here. First, genes encoding the Streptococcus sp. protein G-derived C2 domain (binds to IgG Fc) and Finegoldia magna protein L-derived B1 domain (binds to Igκ light chain) are prepared by PCR amplification. Subsequently, the genes encoding hepatic cell-specific binding domain of hepatitis B virus envelope L protein are replaced by these PCR products. The expression plasmid for this fused gene (encoding GL-fused L protein) can be used to transform Saccharomyces cerevisiae AH22R− cells. To obtain GL-BNC, the transformed yeast cells are disrupted with glass beads, treated with heat, and then subjected to IgG affinity column chromatography followed by size exclusion column chromatography. In addition, GL-BNCs can be fused with liposomes to form GL-virosome. The targeted delivery of GL-BNC and GL-virosome to macrophages can be confirmed by in vitro phagocytosis assays using the murine macrophage cell line RAW264.7.
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Acknowledgments
We would like to address our acknowledgement to Professor Takeshi Arakawa from the University of the Ryukyus for kindly providing the plasmids pCR2.1-SpGB and pET-21d-PpL.
This work was supported in part by JSPS KAKENHI (Grant-in Aid for Scientific Research (S) 16H06314 to S.K.; Grant-in-Aid for Scientific Research (C) 15K07840 to K.T.; Grant-in Aid for JSPS Fellows 18J21249 to H.L.), the Japan Agency for Medical Research and Development (AMED) (19fk0310105h0003 to S.K.), and the Ministry of Education, Culture, Sports, Science and Technology (MEXT) “Dynamic Alliance for Open Innovation Bridging Human, Environment and Materials.”
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Li, H., Somiya, M., Tatematsu, K., Kuroda, S. (2020). Construction of a Macrophage-Targeting Bio-nanocapsule-Based Nanocarrier. In: Jain, K. (eds) Drug Delivery Systems. Methods in Molecular Biology, vol 2059. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9798-5_16
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DOI: https://doi.org/10.1007/978-1-4939-9798-5_16
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