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Production and characterization of a monoclonal antibody specific for Trypanosoma simiae
K M Bosompem1, R K Assoku, V M Nantulya
1Noguchi Memorial Institute for Medical Research, Accra, Ghana.
Monoclonal antibodies (MoAb) were produced against invariant antigens of vector forms of Trypanosoma simiae. X63/AG8.653, NSI/1AG401 and Sp20Ag14 myeloma cells were fused with splenic lymphocytes from BALB/c mice that had been immunized with various preparations of T. simiae procyclics. A T. simiae-specific MoAb [KNS7/14.X (IgG1)] was detected in the hybridoma culture supernatants, which were screened for antibody activity by micro-plate and dot ELISA. Immunofluorescence studies showed that KNS7/14.X stained cytoplasmic antigens in T. simiae procyclics. Proteinase-K digestion and periodate oxidation studies revealed that KNS7/14.X binds to a carbohydrate antigenic determinant in glycoprotein or glycolipid. Cross-reactivity studies using vector forms of T. brucei, T. vivax, T. congolense, T. simiae and T. grayi showed that KNS7/14.X only reacted with T. simiae. Attempts to generate other T. simiae-specific MoAb, using 107-, 75- or 41.7-43.6-kDa peptides selected by western blotting analysis, did not yield positive results.
Monoclonal antibodies (MoAb) were produced against invariant antigens of vector forms of Trypanosoma simiae. X63/AG8.653, NSI/1AG401 and Sp20Ag14 myeloma cells were fused with splenic lymphocytes from BALB/c mice that had been immunized with various preparations of T. simiae procyclics. A T. simiae-specific MoAb [KNS7/14.X (IgG1)] was detected in the hybridoma culture supernatants, which were screened for antibody activity by micro-plate and dot ELISA. Immunofluorescence studies showed that KNS7/14.X stained cytoplasmic antigens in T. simiae procyclics. Proteinase-K digestion and periodate oxidation studies revealed that KNS7/14.X binds to a carbohydrate antigenic determinant in glycoprotein or glycolipid. Cross-reactivity studies using vector forms of T. brucei, T. vivax, T. congolense, T. simiae and T. grayi showed that KNS7/14.X only reacted with T. simiae. Attempts to generate other T. simiae-specific MoAb, using 107-, 75- or 41.7-43.6-kDa peptides selected by western blotting analysis, did not yield positive results.