Secretory production system of bionanocapsules using a stably transfected insect cell line.
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.
Shishido
Takuya
T
Faculty of Engineering, Kobe University, Kobe 657-8501, Japan.
Muraoka
Masaru
M
Ueda
Masakazu
M
Seno
Masaharu
M
Tanizawa
Katsuyuki
K
Kuroda
Shun'ichi
S
Fukuda
Hideki
H
Kondo
Akihiko
A
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
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