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.

ROR1 expression correlated with poor clinical outcome in human ovarian cancer

“…A few studies have suggested several molecular biomarkers of ovarian cancer 26 27 . In our previous research, we have identified several oncogenic biomarkers, constructed related human monoclonal antibody-based drugs and tested their anti-tumor effectiveness 28 29 30 31 . Similarly, we have been searching a novel and valuable marker which associates with ovarian cancer and could be used as a potential candidate for targeted therapy with monoclonal antibodies.…”

Anti‐EBV antibodies: Roles in diagnosis, pathogenesis, and antiviral therapy

“…71 To date, the isolated EBV mAbs mainly target the viral envelope proteins or the viral latent proteins. [72][73][74][75][76][77] Generally, antibodies against the viral envelope proteins interfere the viral entry process, which is known as neutralization, and antibodies targeting viral latent proteins may exert cytotoxic functions. 71,78 Here, we summarized the antiviral potency or complement receptor type2 (CR2), 79 and followingly gp42 binds human leukocyte antigen (HLA) class II to trigger gB conformational change for membrane fusion and viral entry.…”

“…To date, the isolated EBV mAbs mainly target the viral envelope proteins or the viral latent proteins 72–77 . Generally, antibodies against the viral envelope proteins interfere the viral entry process, which is known as neutralization, and antibodies targeting viral latent proteins may exert cytotoxic functions 71,78 .…”

A neutralized human LMP1-IgG inhibits ENKTL growth by suppressing the JAK3/STAT3 signaling pathway

“…Identification of LMP1 expression is drawing attention as a favorable target for ENKTL treatment [ 11 , 12 ]. In our previous researches, we have reported the prognostic characteristics of LMP1 in lymphoma and have generated an anti-LMP1 Fab antibody (LMP1-Fab), exerting potential anti-tumor activity in nasopharyngeal carcinoma (NPC) [ 13 – 18 ]. Because of the remarkable relationships between LMP1 expression and ENKTL properties, an anti-LMP1 antibody should bring positive consequences in ENKTL treatment.…”