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...

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Published in:	Experimental parasitology Vol. 81; no. 4; pp. 536 - 545
Main Authors:	Masake, R.A., Olemoiyoi, O.K., Urakawa, T., Hirumi, H., Majiwa, P.A.O., Wells, C.W., Minja, S.H., Makau, J.M., Nantulya, V.M.
Format:	Journal Article
Language:	English
Published:	San Diego, CA Elsevier Inc 01-12-1995 Elsevier
Subjects:	ADN RECOMBINADO ADN RECOMBINE ANIMAL DEVELOPMENTAL STAGES Animals Antibodies, Monoclonal Antibodies, Protozoan Antibody Specificity ANTICORPS MONOCLONAL ANTICUERPOS MONOCLONALES ANTIGENE ANTIGENOS ANTIGENS Antigens, Protozoan - genetics Antigens, Protozoan - immunology Antigens, Protozoan - isolation & purification BACULOVIRIDAE Biochemistry. Physiology. Immunology. Molecular biology Biological and medical sciences Bombyx - cytology Bombyx - virology Cattle ELISA Enzyme-Linked Immunosorbent Assay - methods EPIMASTIGOTES Escherichia coli ETAPAS DEL DESARROLLO ANIMAL Fundamental and applied biological sciences. Psychology Immunoblotting INFECCIONES POR PROTOZOOS Malaria, Vivax - blood Malaria, Vivax - diagnosis Mice Mice, Inbred BALB C Mice, Inbred C3H Microscopy, Immunoelectron MONOCLONAL ANTIBODIES Nucleopolyhedrovirus - genetics PLASMID VECTORS Protozoa PROTOZOAL INFECTIONS PROTOZOOSE RECOMBINANT DNA Recombinant Fusion Proteins - immunology Spodoptera - cytology Spodoptera - virology STADE DE DEVELOPPEMENT ANIMAL TEST ELISA TRYPANOSOMA VIVAX Trypanosoma vivax - genetics Trypanosoma vivax - immunology Protozoa Protozoal disease Escherichia coli Parasite Parasitosis Monoclonal antibody Baculovirus Gene expression Infection Virus Antigen Baculoviridae Ultrastructure Pathogenic Bacteria Recombinant protein Detection Localization ELISA assay Trypanosomiasis Enterobacteriaceae
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Summary:	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.
Bibliography:	9605624 L72 ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2
ISSN:	0014-4894 1090-2449
DOI:	10.1006/expr.1995.1147