In this article we report the development of an integrated microfluidic system coupled to a screen-printed carbon electrode (SPCE) applied to the quantitative determination of IgG specific antibodies present in serum samples of patients that suffer from Chagas disease. This relevant parasitic infection caused by the hemoflagellate protozoan Trypanosoma cruzi represents a major public health concern in Latin America. In order to perform the detection of mentioned antibodies, SPCE coupled to a microfluidic device was modified by electrodeposition of gold nanoparticles (AuNPs) and functionalized with Trypanosoma cruziproteins from epimastigote membranes. The developed microfluidic immunosensor with immobilized T. cruziproteins on the SPCE surface was successfully applied in the detection of specific IgG anti-T. cruziantibodies, which were allowed to react immunologically with immobilized T. cruziantigen. After that, labelled antibodies were quantified through the addition of horseradish peroxidase (HRP) enzyme-labeled secondary antibodies specific to human IgG, using 4-tert-butylcatechol (4-TBC) as enzymatic mediator. HRP in the presence of hydrogen peroxide (H₂O₂) catalyzes the oxidation of 4-TBC whose back electrochemical reduction was detected on a modified electrode at −100 mV. The calculated detection limit for electrochemical detection was 3.065 ng mL⁻¹ and the intra- and inter-assay coefficients of variation were below 6.95%.