title: Secretion of truncated recombinant rabies virus glycoprotein with preserved antigenic properties using a co-expression system in Hansenula polymorpha.
abstract: Rabies virus infection remains a serious public health threat in the developing world, where cost-concerns make wide-scale public health interventions impractical. The development of novel and inexpensive ELISA diagnostic antigens is critical in early detection and prevention of complications. The transmembrane glycoprotein (G) of rabies virus (RV) contains an external domain capable of inducing the synthesis of anti-rabies, virus-neutralizing antibodies, in infected or immunized hosts. In our study, the external G domain was synthesized and fused in-frame with a polyhistidine-tag coding sequence present in the expression plasmid. Soluble truncated recombinant G was secreted in Hansenula polymorpha (H. polymorpha) using H. polymorpha-derived calnexin (HpCNE1) overproduction and found to be correctly N-glycosylated. The truncated recombinant G was purified from cell culture supernatant by Ni-agarose affinity chromatography and when compared with the full-length glycoprotein, found to be similarly immunogenic in vaccinated rabbits. These results subsequently led us to explore the potential of truncated recombinant G as a diagnostic antigen in ELISA. Our results show that the truncated recombinant G can detect antibodies directed to both whole virion and native glycoprotein. More sophisticated applications of truncated recombinant G would profit from the correctly N-glycosylated and soluble monomer.
authors.affiliation: Life Science and Engineering School, Shaanxi University of Science and Technology, Xi'an 710021, P. R. China.
authors.forename: Weidong
authors.initials: W
authors.lastname: Qian
authors.forename: Frank
authors.initials: F
authors.lastname: Aguilar
authors.forename: Ting
authors.initials: T
authors.lastname: Wang
authors.forename: Bingsheng
authors.initials: B
authors.lastname: Qiu
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D005110:Extracellular Space / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006031:Glycosylation; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D021381:Protein Transport; D011817:Rabbits; D011818:Rabies / Q000276:immunology / Q000517:prevention & control / Q000821:virology; D011819:Rabies Vaccines / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D011820:Rabies virus / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D011819:Rabies Vaccines; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins