Fig 7. ELISA binding assay performed on recombinant glycosomal aldolase expressed in E. coli.
Crude lysate prepared from cultures obtained before induction of protein expression (0 h) and after induction (18 h) were probed. (A) Anti-His IgG or (B) Anti-T. brucei aldolase MAb (Anti-TbALD MAb) was used for probing the presence of expressed aldolase in crude lysate coated on ELISA plate. Anti-His IgG detected all the three recombinant aldolase and Anti-TbALD MAb only detected trypanosomal aldolase (C) Nb474-based homologous sandwich ELISA (C) or (D) Nb474H-Anti-TbALD MAb heterologous sandwich ELISA was used to probe the presence of expressed aldolase in crude lysate. Both homologous (Nb474H-Nb474B) and heterologous (Nb474H-Anti-TbALD MAb) sandwich ELISA detected T. congolense aldolase only. The OD450nm shown on the graphs represent the average value recorded from the duplicate wells.