3.3 Identification of the recombinant ZnT8/GAD65 protein expressed in Sf9 cells by indirect ELISA
Identification of the ZnT8 and GAD65 domains in the recombinant chimera was evaluated by indirect ELISA using ZnT8/GAD65 as the immobilized antigen that interacted with polyclonal rabbit ZnT8 antibodies and monoclonal mouse GAD65 antibodies. As it is shown in Figure 4, both domains were correctly recognized by their specific antibodies, demonstrated by the significant difference of the signals obtained with the specific antibodies with respect to the normal controls. This encouraged us to go ahead with the design of a bridge ELISA for the detection of autoantibodies in individuals at risk or diabetic patients with doubtful classification.