Generation and characterization of a novel recombinant antibody against LMP1-TES1 of Epstein-Barr virus isolated by phage display.

Abstract

Latent Membrane Protein 1 (LMP1) is a primary target for controlling tumorigenesis in Epstein-Barr virus related malignancies; in this study, we aimed to develop a specific antibody against the TES1 domain of the oncogenic LMP1. We screened a full human naïve Fab phage library against TES1 peptide, which consisted of C terminal-activating regions proximal 44 amino acids. After three rounds of panning, enrichment and testing by phage ELISA and further analyzed by DNA sequencing, we selected a phage clone with the highest affinity to LMP1-TES1 and designated it as htesFab. The positive clone was expressed in Escherichia coli and the purified htesFab was characterized for its binding specificity and affinity to LMP1. ELISA, immunofluorescence and FACS analysis confirmed that htesFab could recognize LMP1 TES1 both in vitro and in LMP1 expressing HNE2-LMP1 cells. Furthermore, MTT assay showed that htesFab inhibited the proliferation of HNE2-LMP1 cells in a dose-dependent manner. In summary, this study reported the isolation and characterization of human Fab, which specifically targets the C terminal region/TES1 of LMP1, and has potential to be developed as novel tool for the diagnosis and therapy of Epstein-Barr virus related carcinoma.

• Authors: Dawei Zhang, Yuan Mao, Qing Cao, Lin Xiong, Juan Wen, Renjie Chen, Jin Zhu
• Journal: Viruses (Viruses) Vol. 5 Issue 4 Pg. 1131-42 (Apr 22 2013) ISSN: 1999-4915 [Electronic] Switzerland
• PMID: 23609879 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies
• EBV-associated membrane antigen, Epstein-Barr virus
• Immunoglobulin Fab Fragments
• Peptide Library
• Recombinant Fusion Proteins
• Viral Matrix Proteins

Topics

• Amino Acid Sequence
• Antibodies (immunology)
• Cell Line
• Cell Surface Display Techniques
• Herpesvirus 4, Human (immunology)
• Humans
• Immunoglobulin Fab Fragments (chemistry, genetics, isolation & purification, metabolism)
• Molecular Sequence Data
• Peptide Library
• Protein Binding (immunology)
• Recombinant Fusion Proteins (immunology, metabolism)
• Viral Matrix Proteins (chemistry, immunology, metabolism)