title: Secretory production system of bionanocapsules using a stably transfected insect cell line.
abstract: Bionanocapsules (BNCs) are hollow nanoscale particles composed of L protein of the hepatitis B virus surface antigen that represent specific affinity for human hepatocytes. BNCs can transfer genes and drugs into human hepatocytes efficiently and specifically. BNC can be expressed in yeast cells. In this study, we developed a new L particle production system using a stably transfected insect cell line. For this purpose, we established a host-vector system using the Trichoplusia ni insect cell line. L particles were efficiently secreted by the overexpression of the L protein, which was fused to the secretion signal peptide. The concentration of L particles was reached approximately 1.7 microg/ml in 5 days during cultivation in a serum-free medium without antibiotic selective pressure. The production of L particles was maintained for at least 75 days. The secretory production of L particles facilitated their easy purification by chromatography. Furthermore, it was demonstrated that purified L particles can transfect only human hepatocytes. Therefore, an insect cell expression system is an attractive tool for the production of BNC.
authors.affiliation: Faculty of Engineering, Kobe University, Kobe 657-8501, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Shishido
authors.forename: Masaru
authors.initials: M
authors.lastname: Muraoka
authors.forename: Masakazu
authors.initials: M
authors.lastname: Ueda
authors.forename: Masaharu
authors.initials: M
authors.lastname: Seno
authors.forename: Katsuyuki
authors.initials: K
authors.lastname: Tanizawa
authors.forename: Shun'ichi
authors.initials: S
authors.lastname: Kuroda
authors.forename: Hideki
authors.initials: H
authors.lastname: Fukuda
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kondo
mesh_terms: D000818:Animals; D001516:Bees; D002460:Cell Line; D045744:Cell Line, Tumor; D004330:Drosophila; D018274:Electroporation; D005452:Fluoresceins; D005822:Genetic Vectors; D022781:Hepatocytes; D006801:Humans; D008555:Melitten / Q000737:chemistry; D009036:Moths; D053769:Nanocapsules; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000378:metabolism; D012680:Sensitivity and Specificity; D014162:Transfection; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D005452:Fluoresceins; C096362:L protein, hepatitis B virus; D053769:Nanocapsules; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D008555:Melitten; C007740:fluorexon