Epstein-Barr virus (EBV) has been implicated in the development of many human neoplasias including B lymphomas and nasopharyngeal carcinoma (NPC). The EBV latent membrane protein 1 (LMP-1) has been found to participate in diverse cellular signaling pathways and is essential for virus-induced B-cell immortalization. In order to determine quantitatively the amount of LMP-1 in cells, five monoclonal antibodies (Mabs) specific to LMP-1 were generated. The epitopes recognized by these Mabs were found to cluster within the repeat region between the CTAR1 and CTAR2 domains, corresponding to amino acid positions 254-319 of LMP-1. These Mabs were capable of recognizing LMP-1 proteins of both lymphoid and epithelial origin as revealed by immunoblot, enzyme-linked immunosorbent assay (ELISA) and immunocytofluorescence analysis. A sandwich ELISA for the quantification of LMP-1 has been established using these Mabs. Taken together, our results indicate that the Mabs generated in this study are suitable for the detection of LMP-1 in biomedical research.