Journal: Frontiers in Microbiology

Article Title: Development of oligomannose-coated liposome-based nasal vaccine against human parainfluenza virus type 3

doi: 10.3389/fmicb.2013.00346

Figure Lengend Snippet: Production of full-length HPIV3-HN and its encapsulation into OML. (A) Schematic representation of the expression vector for the wheat germ cell-free system. Full-length HN cDNA was subcloned into either pEU-His or pEU-GST vectors. (B) Expression and solubility of His-HN proteins that were synthesized using the wheat germ cell-free system. The translation reaction mixture (T) was subjected to centrifu-gation followed by the separation of the soluble supernatant (S) and insoluble precipitate (P) fractions. These samples were subjected to SDS-PAGE and visualized with Coomassie Brilliant Blue (CBB) staining. The single asterisk indicates the band of His-HN. (C) Purification of recombinant GST-tagged HPIV3-HN proteins that were synthesized using the wheat germ cell-free system. The translation mixture (T) was subjected to the separation into supernatant (S) and precipitate (P) fractions by centrifugation. GST-HN proteins were purified using glutathione sepharose bead and then resolved by SDS-PAGE and CBB staining. The single asterisk indicates the band of GST-HN. FT, Flow through fraction; E, elute fraction. (D) Schematic of the production of OML-encapsulated His-HN. The single asterisk indicates the band of His-HN. (E) The amount of encapsulated HN was quantitated by SDS-PAGE (right panel). The diameter of OML-HN or control OML were measured by dynamic light scattering particle size analyzer (left panel). The arrow indicates the band of His-HN.

Article Snippet: The particle size of the liposomes was determined using a dynamic light scattering particle size analyzer (BioMedCore Inc., Yokohama, Japan).

Techniques: Encapsulation, Expressing, Plasmid Preparation, Solubility, Synthesized, SDS Page, Staining, Purification, Recombinant, Centrifugation, Control