| Autor: |
Haley DJ; Department of Biomolecular Engineering, University of California Santa Cruz, Santa Cruz, CA 95064, USA., Lanning S; Department of Molecular Cell and Developmental Biology, University of California Santa Cruz, Santa Cruz, CA 95064, USA., Henricson KE; Department of Biomolecular Engineering, University of California Santa Cruz, Santa Cruz, CA 95064, USA., Mardirossian AA; Department of Biomolecular Engineering, University of California Santa Cruz, Santa Cruz, CA 95064, USA., Cirillo I; Department of Biomolecular Engineering, University of California Santa Cruz, Santa Cruz, CA 95064, USA., Rahe MC; Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27607, USA., DuBois RM; Department of Biomolecular Engineering, University of California Santa Cruz, Santa Cruz, CA 95064, USA. |
| Abstrakt: |
Porcine astrovirus 4 (PoAstV4) has been recently associated with respiratory disease in pigs. In order to understand the scope of PoAstV4 infections and to support the development of a vaccine to combat PoAstV4 disease in pigs, we designed and produced a recombinant PoAstV4 capsid spike protein for use as an antigen in serological assays and for potential future use as a vaccine antigen. Structural prediction of the full-length PoAstV4 capsid protein guided the design of the recombinant PoAstV4 capsid spike domain expression plasmid. The recombinant PoAstV4 capsid spike was expressed in Escherichia coli , purified by affinity and size-exclusion chromatography, and its crystal structure was determined at 1.85 Å resolution, enabling structural comparisons to other animal and human astrovirus capsid spike structures. The recombinant PoAstV4 capsid spike protein was also used as an antigen for the successful development of a serological assay to detect PoAstV4 antibodies, demonstrating that the recombinant PoAstV4 capsid spike retains antigenic epitopes found on the native PoAstV4 capsid. These studies lay a foundation for seroprevalence studies and the development of a PoAstV4 vaccine for swine. |
