Purification of the variant antigens of Trypanosoma congolense: a new approach to the isolation of glycoproteins.
We describe a new and rapid method for the isolation and purification of the variant antigens of Trypanosoma congolense. The procedure consists of (a) partial lysis of trypanosomes with dioxane, (b) lectin-affinity-chromatography with Con A-Sepharose, (c) electrophoretic desorption and concomitant separation of Con A-Sepharose-bound glycoproteins in a granulated electrofocusing gel, (d) electrophoretic elution of focused proteins from the granulated gel particles. The efficiency of each step was followed quantitatively by affinity electrophoresis. 73% of the variant antigens originally present in a trypanosomal lysate could be recovered. From 10(10) trypanosomes 2 mg of pure variant antigen were obtained. The variant antigen of the trypanosome clone used exhibits heterogeneity in molecular weight as well as in electric charge.
D000818:Animals; D000954:Antigens, Surface / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D003208:Concanavalin A; D006023:Glycoproteins / Q000276:immunology / Q000302:isolation & purification*; D007525:Isoelectric Focusing; D008970:Molecular Weight; D014345:Trypanosoma / Q000276:immunology*
D000954:Antigens, Surface; D006023:Glycoproteins; D003208:Concanavalin A