Immunological characterization and expression in Escherichia coli and Baculovirus systems of a Trypanosoma vivax antigen detected in the blood of infected animals
A monoclonal antibody (MAb)Tv27 employed in an antigen-detection enzyme immunosorbent assay (Ag-ELISA) for diagnosis of Trypanosoma vivax infection was shown to react with a T. vivax-specific protein of an approximate molecular weight of 10 kDa. This protein is diffusely distributed throughout the c...
| Autores principales: | , , , , , , , , |
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| Formato: | Journal Article |
| Lenguaje: | Inglés |
| Publicado: |
Elsevier
1995
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| Materias: | |
| Acceso en línea: | https://hdl.handle.net/10568/28608 |
| _version_ | 1855520675842752512 |
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| author | Masake, R.A. Ole-MoiYoi, O.K. Urakawa, T. Hirumi, H. Majiwa, Phelix A.O. Wells, C.W. Minja, S.H. Makau, J.M. Nantulya, V.M. |
| author_browse | Hirumi, H. Majiwa, Phelix A.O. Makau, J.M. Masake, R.A. Minja, S.H. Nantulya, V.M. Ole-MoiYoi, O.K. Urakawa, T. Wells, C.W. |
| author_facet | Masake, R.A. Ole-MoiYoi, O.K. Urakawa, T. Hirumi, H. Majiwa, Phelix A.O. Wells, C.W. Minja, S.H. Makau, J.M. Nantulya, V.M. |
| author_sort | Masake, R.A. |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | A monoclonal antibody (MAb)Tv27 employed in an antigen-detection enzyme immunosorbent assay (Ag-ELISA) for diagnosis of Trypanosoma vivax infection was shown to react with a T. vivax-specific protein of an approximate molecular weight of 10 kDa. This protein is diffusely distributed throughout the cytosol and nucleus of metacyclic forms, bloodstream forms, and procyclic-like elongated trypomastigotes, but is not detectable in epimastigotes of T. vivax. The T. vivax-specific antigen prepared from parasite lysates appeared to be of lower molecular mass than the form expressed in either Escherichia coli or in baculovirus-infected silkworm insect cells. In the recombinant baculovirus-infected cells, the protein was expressed mostly as an 18-kDa peptide with less abundant forms of 13 and 12 kDa, while the protein expressed in E. coli was approximately 14 kDa. Both the low- and higher-molecular-weight proteins are recognized by the MAb Tv27 in Western blots and in Ag-ELISA. Although the crude preparations of the protein produced by the insect cells are labile when kept for more than 2 hr at 24°C, they retained reactivity at temperatures below 4°C for several weeks. The proteins expressed in both the insect cells and E. coli captured anti-T. vivax antibodies in sera prepared from trypanosome-infected animals. Since the recombinant protein expressed in the baculovirus-infected cells is available in large homogenous quantities, it would serve as a positive control in Ag-ELISA and is also usable for antibody detection assays. |
| format | Journal Article |
| id | CGSpace28608 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 1995 |
| publishDateRange | 1995 |
| publishDateSort | 1995 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | CGSpace286082024-05-01T08:16:21Z Immunological characterization and expression in Escherichia coli and Baculovirus systems of a Trypanosoma vivax antigen detected in the blood of infected animals Masake, R.A. Ole-MoiYoi, O.K. Urakawa, T. Hirumi, H. Majiwa, Phelix A.O. Wells, C.W. Minja, S.H. Makau, J.M. Nantulya, V.M. trypanosomiasis trypanosoma vivax escherichia coli immunology disease control glossinidae antibodies animal diseases antigens monoclonal antibodies A monoclonal antibody (MAb)Tv27 employed in an antigen-detection enzyme immunosorbent assay (Ag-ELISA) for diagnosis of Trypanosoma vivax infection was shown to react with a T. vivax-specific protein of an approximate molecular weight of 10 kDa. This protein is diffusely distributed throughout the cytosol and nucleus of metacyclic forms, bloodstream forms, and procyclic-like elongated trypomastigotes, but is not detectable in epimastigotes of T. vivax. The T. vivax-specific antigen prepared from parasite lysates appeared to be of lower molecular mass than the form expressed in either Escherichia coli or in baculovirus-infected silkworm insect cells. In the recombinant baculovirus-infected cells, the protein was expressed mostly as an 18-kDa peptide with less abundant forms of 13 and 12 kDa, while the protein expressed in E. coli was approximately 14 kDa. Both the low- and higher-molecular-weight proteins are recognized by the MAb Tv27 in Western blots and in Ag-ELISA. Although the crude preparations of the protein produced by the insect cells are labile when kept for more than 2 hr at 24°C, they retained reactivity at temperatures below 4°C for several weeks. The proteins expressed in both the insect cells and E. coli captured anti-T. vivax antibodies in sera prepared from trypanosome-infected animals. Since the recombinant protein expressed in the baculovirus-infected cells is available in large homogenous quantities, it would serve as a positive control in Ag-ELISA and is also usable for antibody detection assays. 1995-12 2013-05-06T07:00:59Z 2013-05-06T07:00:59Z Journal Article https://hdl.handle.net/10568/28608 en Limited Access Elsevier Experimental Parasitology;81: 536-545 |
| spellingShingle | trypanosomiasis trypanosoma vivax escherichia coli immunology disease control glossinidae antibodies animal diseases antigens monoclonal antibodies Masake, R.A. Ole-MoiYoi, O.K. Urakawa, T. Hirumi, H. Majiwa, Phelix A.O. Wells, C.W. Minja, S.H. Makau, J.M. Nantulya, V.M. Immunological characterization and expression in Escherichia coli and Baculovirus systems of a Trypanosoma vivax antigen detected in the blood of infected animals |
| title | Immunological characterization and expression in Escherichia coli and Baculovirus systems of a Trypanosoma vivax antigen detected in the blood of infected animals |
| title_full | Immunological characterization and expression in Escherichia coli and Baculovirus systems of a Trypanosoma vivax antigen detected in the blood of infected animals |
| title_fullStr | Immunological characterization and expression in Escherichia coli and Baculovirus systems of a Trypanosoma vivax antigen detected in the blood of infected animals |
| title_full_unstemmed | Immunological characterization and expression in Escherichia coli and Baculovirus systems of a Trypanosoma vivax antigen detected in the blood of infected animals |
| title_short | Immunological characterization and expression in Escherichia coli and Baculovirus systems of a Trypanosoma vivax antigen detected in the blood of infected animals |
| title_sort | immunological characterization and expression in escherichia coli and baculovirus systems of a trypanosoma vivax antigen detected in the blood of infected animals |
| topic | trypanosomiasis trypanosoma vivax escherichia coli immunology disease control glossinidae antibodies animal diseases antigens monoclonal antibodies |
| url | https://hdl.handle.net/10568/28608 |
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