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.
| Main Authors: | , , , , , , , , |
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| Format: | Journal Article biblioteca |
| Language: | English |
| Published: |
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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