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Immunogenic and antigenic properties of recombinant soluble glycoprotein of rabies virus.
- Source :
-
Veterinary microbiology [Vet Microbiol] 2005 Jul 01; Vol. 108 (3-4), pp. 207-14. - Publication Year :
- 2005
-
Abstract
- Rabies virus glycoprotein is a type I transmembrane protein exposed on the surface on the mature virus particle that induces virus neutralizing antibodies. In the present study, 60 amino acid C-terminal hydrophobic anchor (transmembrane) and cytoplasmic domains of glycoprotein were deleted from full-length glycoprotein and fused with polyhistidine tag. The N-terminal viral signal peptide was also replaced with CD33 signal peptide for efficient secretion in mammalian cells. Following transfection of Madin Darby bovine kidney (MDBK) cells with plasmid encoding this soluble form of glycoprotein, polyclonal populations of stably transfected resistant cells were obtained after G418 selection. The protein was expressed as a glycosylated protein and secreted outside the cells utilizing N-terminal CD33 signal peptide. The secreted soluble glycoprotein was purified from cell culture supernatant by Ni--agarose affinity chromatography utilizing C-terminal polyhistidine tag. Like full-length glycoprotein, the expressed recombinant soluble glycoprotein was found to be immunogenic when injected in rabbits. In this study, we have assessed the potential of recombinant soluble glycoprotein as diagnostic antigen in ELISA and found that this recombinant protein can be used as diagnostic antigen in ELISA for detecting anti-glycoprotein antibodies in immunized host.
- Subjects :
- Animals
Antibodies, Viral blood
Antigens, Viral genetics
Blotting, Western
Chromatography, Affinity
DNA, Viral chemistry
DNA, Viral genetics
Dog Diseases virology
Dogs
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay
Escherichia coli genetics
Glycoproteins genetics
Immunization
Polymerase Chain Reaction
Rabbits
Rabies virology
Rabies virus genetics
Recombinant Proteins genetics
Recombinant Proteins immunology
Sensitivity and Specificity
Transfection
Viral Envelope Proteins genetics
Antigens, Viral immunology
Glycoproteins immunology
Rabies virus immunology
Viral Envelope Proteins immunology
Subjects
Details
- Language :
- English
- ISSN :
- 0378-1135
- Volume :
- 108
- Issue :
- 3-4
- Database :
- MEDLINE
- Journal :
- Veterinary microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 15916870
- Full Text :
- https://doi.org/10.1016/j.vetmic.2005.04.007