Construction and characterization of monoclonal antibodies specific to Epstein-Barr virus latent membrane protein 1.
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.
Fang
Chih-Yeu
CY
Institute of Molecular Medicine, National Tsing Hua University, Hsin-chu 300, Taiwan, ROC.
Chang
Yu-Sun
YS
Chow
Kai-Ping
KP
Yu
Jau-Song
JS
Chang
Hwan-You
HY
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D002460:Cell Line; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D018985:Epitopes, B-Lymphocyte / Q000276:immunology; D005260:Female; D005455:Fluorescent Antibody Technique; D005982:Glutathione Transferase / Q000235:genetics / Q000276:immunology; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006801:Humans; D015151:Immunoblotting; D007132:Immunoglobulin Isotypes; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D014162:Transfection; D014763:Viral Matrix Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D017931:DNA Primers; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D018985:Epitopes, B-Lymphocyte; D007132:Immunoglobulin Isotypes; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins; D005982:Glutathione Transferase