Novel biotechnological platform based on baculovirus occlusion bodies carrying Babesia bovis small antigenic peptides for the design of a diagnostic enzyme-linked immunosorbent assay (ELISA)
Baculoviruses are large DNA virus of insects principally employed in recombinant protein expression. Its ability to form occlusion bodies (OBs), which are composed mainly of polyhedrin protein (POLH), makes them biotechnologically attractive, as these crystals (polyhedra) can incorporate foreign pep...
| Autores principales: | , , , , , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
| Publicado: |
Springer
2018
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| Materias: | |
| Acceso en línea: | https://link.springer.com/article/10.1007%2Fs00253-017-8662-1 http://hdl.handle.net/20.500.12123/3626 https://doi.org/10.1007/s00253-017-8662-1 |
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| author | Lopez, Maria Gabriela Pallares, H.M. Alfonso, Victoria Carmona, S. J. Farber, Marisa Diana Taboga, Oscar Alberto Wilkowsky, Silvina Elizabeth |
| author_browse | Alfonso, Victoria Carmona, S. J. Farber, Marisa Diana Lopez, Maria Gabriela Pallares, H.M. Taboga, Oscar Alberto Wilkowsky, Silvina Elizabeth |
| author_facet | Lopez, Maria Gabriela Pallares, H.M. Alfonso, Victoria Carmona, S. J. Farber, Marisa Diana Taboga, Oscar Alberto Wilkowsky, Silvina Elizabeth |
| author_sort | Lopez, Maria Gabriela |
| collection | INTA Digital |
| description | Baculoviruses are large DNA virus of insects principally employed in recombinant protein expression. Its ability to form occlusion bodies (OBs), which are composed mainly of polyhedrin protein (POLH), makes them biotechnologically attractive, as these crystals (polyhedra) can incorporate foreign peptides and can be easily isolated. On the other hand, peptide microarrays allow rapid and inexpensive high-throughput serological screening of new candidates to be incorporated to OBs. To integrate these 2 biotechnological approaches, we worked on Babesia bovis, one of the causative agents of bovine babesiosis. Current molecular diagnosis of infection with B. bovis includes enzyme-linked immunosorbent assay (ELISA) techniques, which use merozoite lysate obtained from infected bovine erythrocytes. However, it is important to produce recombinant antigens that replace the use of crude antigens. Here, we describe a new biotechnological platform for the design of indirect ELISAs based on 5 antigenic peptides of 15 amino acid residues of B. bovis (ApBb), selected from a peptide microarray and expressed as a fusion to POLH. An Sf9POLHE44G packaging cell line infected with recombinant baculoviruses carrying POLH-ApBb fusions yielded higher levels of chimeric polyhedra, highlighting the advantage of a trans-contribution of a mutant copy of polyhedrin. Finally, the use of dissolved recombinant polyhedra as antigens was successful in an ELISA assay, as B. bovis-positive sera recognized the fusion POLH-ApBb. Thus, the use of this platform resulted in a promising alternative for molecular diagnosis of relevant infectious diseases. |
| format | Artículo |
| id | INTA3626 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2018 |
| publishDateRange | 2018 |
| publishDateSort | 2018 |
| publisher | Springer |
| publisherStr | Springer |
| record_format | dspace |
| spelling | INTA36262018-10-18T13:00:30Z Novel biotechnological platform based on baculovirus occlusion bodies carrying Babesia bovis small antigenic peptides for the design of a diagnostic enzyme-linked immunosorbent assay (ELISA) Lopez, Maria Gabriela Pallares, H.M. Alfonso, Victoria Carmona, S. J. Farber, Marisa Diana Taboga, Oscar Alberto Wilkowsky, Silvina Elizabeth Baculovirus Elisa Babesia bovis Péptidos Antígenos Virales Técnicas Inmunoenzimáticas Peptides Viral Antigens Immunoenzyme Techniques Baculoviruses are large DNA virus of insects principally employed in recombinant protein expression. Its ability to form occlusion bodies (OBs), which are composed mainly of polyhedrin protein (POLH), makes them biotechnologically attractive, as these crystals (polyhedra) can incorporate foreign peptides and can be easily isolated. On the other hand, peptide microarrays allow rapid and inexpensive high-throughput serological screening of new candidates to be incorporated to OBs. To integrate these 2 biotechnological approaches, we worked on Babesia bovis, one of the causative agents of bovine babesiosis. Current molecular diagnosis of infection with B. bovis includes enzyme-linked immunosorbent assay (ELISA) techniques, which use merozoite lysate obtained from infected bovine erythrocytes. However, it is important to produce recombinant antigens that replace the use of crude antigens. Here, we describe a new biotechnological platform for the design of indirect ELISAs based on 5 antigenic peptides of 15 amino acid residues of B. bovis (ApBb), selected from a peptide microarray and expressed as a fusion to POLH. An Sf9POLHE44G packaging cell line infected with recombinant baculoviruses carrying POLH-ApBb fusions yielded higher levels of chimeric polyhedra, highlighting the advantage of a trans-contribution of a mutant copy of polyhedrin. Finally, the use of dissolved recombinant polyhedra as antigens was successful in an ELISA assay, as B. bovis-positive sera recognized the fusion POLH-ApBb. Thus, the use of this platform resulted in a promising alternative for molecular diagnosis of relevant infectious diseases. Instituto de Biotecnología Fil: Lopez, Maria Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Pallares, H. M. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Alfonso, Victoria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Carmona, S. J. University of Lausanne. Ludwig Cancer Research Center. Department of Fundamental Oncology; Suiza. Swiss Institute of Bioinformatics (SIB); Suiza Fil: Farber, Marisa Diana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Wilkowsky, Silvina Elizabeth. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina 2018-10-18T12:56:45Z 2018-10-18T12:56:45Z 2018-01 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion https://link.springer.com/article/10.1007%2Fs00253-017-8662-1 http://hdl.handle.net/20.500.12123/3626 1432-0614 https://doi.org/10.1007/s00253-017-8662-1 eng info:eu-repograntAgreement/INTA/PNBIO/1131032/AR./Desarrollo de herramientas biotecnológicas para la prevención y el control de enfermedades pecuarias: vacunas, diagnóstico y eIdemiología molecular. info:eu-repo/semantics/restrictedAccess application/pdf Springer Applied Microbiology and Biotechnology 102 (2): 885–896 (enero 2018) |
| spellingShingle | Baculovirus Elisa Babesia bovis Péptidos Antígenos Virales Técnicas Inmunoenzimáticas Peptides Viral Antigens Immunoenzyme Techniques Lopez, Maria Gabriela Pallares, H.M. Alfonso, Victoria Carmona, S. J. Farber, Marisa Diana Taboga, Oscar Alberto Wilkowsky, Silvina Elizabeth Novel biotechnological platform based on baculovirus occlusion bodies carrying Babesia bovis small antigenic peptides for the design of a diagnostic enzyme-linked immunosorbent assay (ELISA) |
| title | Novel biotechnological platform based on baculovirus occlusion bodies carrying Babesia bovis small antigenic peptides for the design of a diagnostic enzyme-linked immunosorbent assay (ELISA) |
| title_full | Novel biotechnological platform based on baculovirus occlusion bodies carrying Babesia bovis small antigenic peptides for the design of a diagnostic enzyme-linked immunosorbent assay (ELISA) |
| title_fullStr | Novel biotechnological platform based on baculovirus occlusion bodies carrying Babesia bovis small antigenic peptides for the design of a diagnostic enzyme-linked immunosorbent assay (ELISA) |
| title_full_unstemmed | Novel biotechnological platform based on baculovirus occlusion bodies carrying Babesia bovis small antigenic peptides for the design of a diagnostic enzyme-linked immunosorbent assay (ELISA) |
| title_short | Novel biotechnological platform based on baculovirus occlusion bodies carrying Babesia bovis small antigenic peptides for the design of a diagnostic enzyme-linked immunosorbent assay (ELISA) |
| title_sort | novel biotechnological platform based on baculovirus occlusion bodies carrying babesia bovis small antigenic peptides for the design of a diagnostic enzyme linked immunosorbent assay elisa |
| topic | Baculovirus Elisa Babesia bovis Péptidos Antígenos Virales Técnicas Inmunoenzimáticas Peptides Viral Antigens Immunoenzyme Techniques |
| url | https://link.springer.com/article/10.1007%2Fs00253-017-8662-1 http://hdl.handle.net/20.500.12123/3626 https://doi.org/10.1007/s00253-017-8662-1 |
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