title: Protein-encapsulated bio-nanocapsules production with ER membrane localization sequences.
abstract: Bio-nanocapsules (BNCs) are hollow nanoparticles composed of the L protein of hepatitis B virus (HBV) surface antigen (HBsAg), which can specifically introduce genes and drugs into various kinds of target cells. Although the classic electroporation method has typically been used to introduce highly charged molecules such as DNA, it is rarely adopted for proteins due to its very low efficiency. In this study, a novel approach to the preparation of BNC was established whereby a target protein was pre-encapsulated during the course of nanoparticle formation. Briefly, because of the process of BNC formation in a budding manner on the endoplasmic reticulum (ER) membrane, the association of target proteins to the ER membrane with lipidation sequences (ER membrane localization sequences) could directly generate protein-encapsulating BNC in collaboration with co-expression of the L proteins. Since the membrane-localized proteins are automatically enveloped into BNCs during the budding event, this method can be protect the proteins and BNCs from damage caused by electroporation and obviate the need for laborious consideration to study the optimal conditions for protein encapsulation. This approach would be a useful method for encapsulating therapeutic candidate proteins into BNCs.
authors.affiliation: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe 657-8501, Japan.
authors.forename: Yuya
authors.initials: Y
authors.lastname: Nishimura
authors.forename: Takuya
authors.initials: T
authors.lastname: Shishido
authors.forename: Jun
authors.initials: J
authors.lastname: Ishii
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Tanaka
authors.forename: Chiaki
authors.initials: C
authors.lastname: Ogino
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kondo
mesh_terms: D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D011905:Genes, ras; D006514:Hepatitis B Surface Antigens / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007313:Insecta; D042541:Intracellular Space / Q000378:metabolism; D050356:Lipid Metabolism; D053769:Nanocapsules / Q000737:chemistry*; D010316:Particle Size; D010957:Plasmids; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D006514:Hepatitis B Surface Antigens; C096362:L protein, hepatitis B virus; D053769:Nanocapsules; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins