Monoclonal antibodies (mAbs) were raised against hepatitis B virus core produced by a recombinant clone of Escherichia coli (rHBc). The three mAbs recognized rHBc by Western blot, suggesting that they reacted with non-conformational epitopes. Competition experiments between mAbs and human anti-HBc sera confirmed the existence of an immunodominant HBc epitope within the viral antigen. A monoclonal competition enzyme immunoassay using an IgM mAb conjugated to biotin and streptavidin-peroxidase as the detection system yielded 99% sensitivity and 100% specificity, when compared to other commercial assays.