Figure 3.
Western blotting analyses of purified TGRA8. A) Reactivity of purified TGRA8 with human sera. Purified TGRA8 was analyzed on the SDS-PAGE, transferred onto the nitrocellulose membrane and probed with the acute and chronic pooled sera from pregnant women. The purified antigen strongly reacted with IgG and IgM antibodies in acute sera, while no specific band can be observed on the blot probed with chronic sera,. The protein band of about 54 kDa was probably attributed to the dimmer of TGRA8. B) Evaluation of immunogenicity of TGRA8. One Rabbit was injected intramuscularly with 100 μg of TGRA8 three times with two weeks intervals. Two weeks after the last injection rabbit serum sample was obtained. Tachyzoites of T.gondii RH strain (5×106 tachyzoites) were analyzed by SDS-PAGE, transferred onto nitrocellulose membrane and probed with the serum samples from the TGRA8-immunized and adjuvant-injected rabbits. The serum from the TGRA8-immunized rabbit recognized a single protein band of 35 kDa, the expected size of the native GRA8 antigen, while the serum from the adjuvant-injected rabbit failed to react with any specific protein of RH tachyzoites