The Use of Chimeric Virus-like Particles Harbouring a Segment of Hantavirus Gc Glycoprotein to Generate a Broadly-Reactive Hantavirus-Specific Monoclonal Antibody
Abstract
:1. Introduction
2. Results
2.1. Production of Recombinant Full-Length PUUV-Gc Protein in Yeast S. cerevisiae and Generation of Monoclonal Antibodies
2.2. Expression, Purification and Electron Microscopy Characterization of Chimeric VLPs Harbouring PUUV-Gc Protein Segment
2.3. Antigenic Properties of Chimeric VLPs Harbouring the Gc99 Protein Segment
2.4. Immunogenic Properties of Chimeric VLPs
2.5. Generation of Monoclonal Antibodies against Hantavirus Gc Segment Displayed on VLPs
2.6. Mapping of the MAb #10B8 Epitope
2.7. The Reactivity of the MAb #10B8 with Hantavirus-Infected Cells
2.8. Neutralization Test with the MAb #10B8
3. Discussion
4. Experimental Section
4.1. Monoclonal Antibodies and Polyclonal Antisera
4.2. Generation of an Expression Plasmid Encoding Full-Length PUUV Gc Glycoprotein
4.3. Expression and Purification of Recombinant Full-Length Hantavirus Gc and N Proteins
4.4. Generation of Expression Plasmids Encoding a VP1 Gc Fusion Protein
4.5. Expression and Characterization of Chimeric VP1 Protein Harbouring PUUV Gc Protein Insert
4.6. SDS-PAGE and Western Blot Analysis
4.7. Immunization of Mice and Generation of Monoclonal Antibodies
4.8. Indirect ELISA
4.9. Indirect Immunofluorescence Assay
4.10. Hantavirus Neutralization Test
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References and Notes
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Zvirbliene, A.; Kucinskaite-Kodze, I.; Razanskiene, A.; Petraityte-Burneikiene, R.; Klempa, B.; Ulrich, R.G.; Gedvilaite, A. The Use of Chimeric Virus-like Particles Harbouring a Segment of Hantavirus Gc Glycoprotein to Generate a Broadly-Reactive Hantavirus-Specific Monoclonal Antibody. Viruses 2014, 6, 640-660. https://doi.org/10.3390/v6020640
Zvirbliene A, Kucinskaite-Kodze I, Razanskiene A, Petraityte-Burneikiene R, Klempa B, Ulrich RG, Gedvilaite A. The Use of Chimeric Virus-like Particles Harbouring a Segment of Hantavirus Gc Glycoprotein to Generate a Broadly-Reactive Hantavirus-Specific Monoclonal Antibody. Viruses. 2014; 6(2):640-660. https://doi.org/10.3390/v6020640
Chicago/Turabian StyleZvirbliene, Aurelija, Indre Kucinskaite-Kodze, Ausra Razanskiene, Rasa Petraityte-Burneikiene, Boris Klempa, Rainer G. Ulrich, and Alma Gedvilaite. 2014. "The Use of Chimeric Virus-like Particles Harbouring a Segment of Hantavirus Gc Glycoprotein to Generate a Broadly-Reactive Hantavirus-Specific Monoclonal Antibody" Viruses 6, no. 2: 640-660. https://doi.org/10.3390/v6020640