Production and characterization of a monoclonal antibody specific for Trypanosoma simiae.
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.
Bosompem
K M
KM
Noguchi Memorial Institute for Medical Research, Accra, Ghana.
Assoku
R K
RK
Nantulya
V M
VM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000145:classification; D000913:Antibodies, Protozoan / Q000096:biosynthesis* / Q000145:classification; D000918:Antibody Specificity; D000953:Antigens, Protozoan / Q000737:chemistry / Q000276:immunology; D003429:Cross Reactions; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D051381:Rats; D017207:Rats, Sprague-Dawley; D013552:Swine / Q000469:parasitology; D014345:Trypanosoma / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan