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 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.
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Format: | Journal Article biblioteca |
Language: | English |
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Elsevier
1995-12
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Subjects: | trypanosomiasis, trypanosoma vivax, escherichia coli, immunology, disease control, glossinidae, antibodies, animal diseases, antigens, monoclonal antibodies, |
Online Access: | https://hdl.handle.net/10568/28608 https://doi.org/10.1006/expr.1995.1147 |
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dig-cgspace-10568-286082023-12-08T19:36:04Z 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, P.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 Experimental Parasitology;81: 536-545 0014-4894 https://hdl.handle.net/10568/28608 https://doi.org/10.1006/expr.1995.1147 en Copyrighted; all rights reserved Limited Access p. 536-545 Elsevier Experimental Parasitology |
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trypanosomiasis trypanosoma vivax escherichia coli immunology disease control glossinidae antibodies animal diseases antigens monoclonal antibodies trypanosomiasis trypanosoma vivax escherichia coli immunology disease control glossinidae antibodies animal diseases antigens monoclonal antibodies |
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trypanosomiasis trypanosoma vivax escherichia coli immunology disease control glossinidae antibodies animal diseases antigens monoclonal antibodies 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, P.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 |
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 |
topic_facet |
trypanosomiasis trypanosoma vivax escherichia coli immunology disease control glossinidae antibodies animal diseases antigens monoclonal antibodies |
author |
Masake, R.A. Ole-MoiYoi, O.K. Urakawa, T. Hirumi, H. Majiwa, P.A.O. Wells, C.W. Minja, S.H. Makau, J.M. Nantulya, V.M. |
author_facet |
Masake, R.A. Ole-MoiYoi, O.K. Urakawa, T. Hirumi, H. Majiwa, P.A.O. Wells, C.W. Minja, S.H. Makau, J.M. Nantulya, V.M. |
author_sort |
Masake, R.A. |
title |
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_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_sort |
immunological characterization and expression in escherichia coli and baculovirus systems of a trypanosoma vivax antigen detected in the blood of infected animals |
publisher |
Elsevier |
publishDate |
1995-12 |
url |
https://hdl.handle.net/10568/28608 https://doi.org/10.1006/expr.1995.1147 |
work_keys_str_mv |
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