1. mAb CZP-315.D9: An Antirecombinant Cruzipain Monoclonal Antibody That Specifically Labels the Reservosomes of Trypanosoma cruzi Epimastigotes
- Author
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Iriane Eger, Maurilio J. Soares, Lia Carolina Soares Medeiros, and Cassiano Martin Batista
- Subjects
Chagas disease ,Article Subject ,medicine.drug_class ,Trypanosoma cruzi ,Protozoan Proteins ,lcsh:Medicine ,Cruzipain ,Immunofluorescence ,Monoclonal antibody ,General Biochemistry, Genetics and Molecular Biology ,law.invention ,Mice ,Microscopy, Electron, Transmission ,law ,parasitic diseases ,medicine ,Animals ,Humans ,Chagas Disease ,Organelles ,General Immunology and Microbiology ,medicine.diagnostic_test ,biology ,lcsh:R ,Antibodies, Monoclonal ,Cell Differentiation ,General Medicine ,biology.organism_classification ,medicine.disease ,Molecular biology ,Endocytosis ,Cysteine Endopeptidases ,Monoclonal ,biology.protein ,Recombinant DNA ,Antibody ,Lysosomes ,Research Article - Abstract
Reservosomes are large round vesicles located at the posterior end of epimastigote forms of the protozoanTrypanosoma cruzi, the etiological agent of Chagas disease. They are the specific end organelles of the endocytosis pathway ofT. cruzi, and they play key roles in nutrient uptake and cell differentiation. These lysosome-like organelles accumulate ingested macromolecules and contain large amounts of a major cysteine proteinase (cruzipain or GP57/51 protein). Aim of this study was to produce a monoclonal antibody (mAb) against a recombinantT. cruzicruzipain (TcCruzipain) that specifically labels the reservosomes. BALB/c mice were immunized with purified recombinant TcCruzipain to obtain the mAb. After fusion of isolated splenocytes with myeloma cells and screening, a mAb was obtained by limiting dilution and characterized by capture ELISA. We report here the production of a kappa-positive monoclonal IgG antibody (mAb CZP-315.D9) that recognizes recombinant TcCruzipain. This mAb binds preferentially to a protein with a molecular weight of about 50 kDa on western blots and specifically labels reservosomes by immunofluorescence and transmission electron microscopy. The monoclonal CZP-315.D9 constitutes a potentially powerful marker for use in studies on the function of reservosomes ofT. cruzi.
- Published
- 2014