Use of Protein A as the Primary Layer in Fluorescent Microsphere Technology

摘要

Six unique monoclonal antibodies (mAb) to the galactose-binding adherence lectinof Entamoeba histolytica were assessed for their abilities to bind protein A, using BIAcore. Of the six antibodies tested, mAb H85, specific for pathogenic strains, showed the greatest degree of binding to Protein A. Therefore, protein A was used as a surface layer on microspheres to which mAbs H85 and 3F4 were attached. Conversion of this method to microsphere agglutination and enzyme linked immunabsorbent assay (ELISA) formats proved successful. Instrumented reads of agglutination reactions in the ELISA format achieved a lower limit of detection equivalent to 500 amoeba when performed on solubilized amebic antigen. Protein A, Fluorescence, Epitope, Adherence lectin, Microsphere, Monoclonal antibody.