Recombinant dengue virus type 2 envelope/hepatitis B surface antigen hybrid protein expressed in Pichia pastoris can function as a bivalent immunogen.
A truncated version of the dengue virus type 2 envelope protein (Den2E) encoding the first 395 amino acid (aa) residues, and Den2E fused in-frame with the full-length 226-aa hepatitis B surface antigen (Den2E-HBsAg) protein were expressed in the methylotrophic yeast, Pichia pastoris. Both the recombinant proteins showed evidence of the capacity to form high molecular weight aggregates. Electron microscopic analysis of the purified proteins showed that while Den2E displayed an amorphous morphology, Den2E-HBsAg existed as well-structured virus-like particles (VLPs). Using immuno-gold electron microscopy, these VLPs were demonstrated to contain both components of the Den2E-HBsAg hybrid protein. Seroanalysis showed that the hybrid VLPs could function in vivo as bivalent immunogens, which could elicit immune responses directed against both components of the hybrid protein, as evidenced by ELISA, immunoprecipitation and immunofluorescence data.
Bisht
Himani
H
International Centre for Genetic Engineering and Biotechnology, RGP Laboratory, PO Box 10504, Aruna Asaf Ali Marg, New Delhi 110067, India.
Chugh
Dipti A
DA
Raje
Manoj
M
Swaminathan
S Swaminathan
SS
Khanna
Navin
N
D000818:Animals; D000911:Antibodies, Monoclonal; D015870:Gene Expression; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D010843:Pichia; D011946:Receptors, Antigen / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D014614:Vaccines, Synthetic / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; C051821:E-glycoprotein, Dengue virus type 2; D011946:Receptors, Antigen; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; C030500:hepatitis B surface antigen receptor