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Construction and characterization of monoclonal antibodies specific to Epstein-Barr virus latent membrane protein 1
Journal article   Peer reviewed

Construction and characterization of monoclonal antibodies specific to Epstein-Barr virus latent membrane protein 1

Chih-Yeu Fang, Yu-Sun Chang, Kai-Ping Chow, Jau-Song Yu and Hwan-You Chang
Journal of Immunological Methods, Vol.287(1-2), pp.21-30
04/2004

Abstract

5-bromo-4-chloro-3- indolyl phosphate BCIP DMEM Dulbecco's modified Eagle's medium EBV ELISA Enzyme-linked immunosorbent assay Epstein-Barr virus FBS Fetal bovine serum FITC Fluorescein isothiocyanate LMP-1 Monoclonal antibody Quantitative ELISA
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. © 2004 Elsevier B.V. All rights reserved.

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