DEVELOPENZYMET AND QUALITY ASSESSMENT MOUSE OF ANTI-HUMAN IgG ANTIBODYHORSERADISH PEROXIDASE CONJUGATES USING FOR ELISA TECHNIQUE TO DETECT HUMAN ANTIBOBY

Abstract

HRP (Horseradish peroxidase) has been known as an enzyme extracted from horseradish roots (Amoracia Rusticana). HRP plays an important role as a marker which most used as a common method of conjugations due to the relatively cheap, purified, fast signaling and highly specific enzyme. Furthermore, HRP involves small sizes and molecular weights having less obstructive space in associated with antigen or antibody. In immunological tests, such as ELISA, secondary antibody – horseradish peroxidase conjugates has been applied to increase specificity of enzyme in ELISA test such as anti-IgG, anti-IgM. Primary mice antibody-conjugated enzymes which also practiced for any serum sample performing any anti-antigen functions were observed. This project objects to conduct an immune response in mice with IgG antibodies which were purified from the serum of umbilical cord blood injected in different doses of 20 μg, 40 μg and 60 μg per dose of vaccine. After purified mouse antibody concentrations obtained the highest 0.495 mg per ml. The antibody anti-mouse antibody has been added in associated with the HRP enzymes. In conclusion, the antibody-antigen interaction was examined by direct and indirect ELISA and the level of community was in concentration of 0.125 mg/ml.

Key words: HRP, horseradish root, immune response, IgG, IgM, ELISA, serum.