The enzyme-linked immunosorbent assay (ELISA) is a frequently used analytical immunochemistry assay for detection and quantification of different molecules such as proteins and antibodies. This work described the optimization of non competitive indirect ELISA for the quantification of total IgG antibodies against ovalbumin protein in the Balb/c mice serum after immunization of animals with ovalbumin as an immunogen. The evaluation of the sensitivity of the test with regard to the different dilutions and concentrations used makes it possible to set the optimal concentrations of the different parameters, namely the dilutions of the anti-OVA mice immune serum, of the goat immunoglobulins labeled with horseradish peroxidase (conjugate) and concentrations of ovalbumin antigen. Our results show that the optimization of this ELISA test made it possible to set the concentration of the antigen: OVA at 5µg/ml when the dilutions in anti-OVA mice immune serum and in conjugate are respectively 1/300 and 1/2000. The possibilities of application of immunochemical analysis techniques such as ELISA in medical field and in food industry are very wide, whether to advantageously replace (specificity, simplicity) other analysis techniques, or to make up for a lack of analysis resources.