Generation and Characterization of a Novel Recombinant Antibody against LMP1-TES1 of Epstein-Barr Virus Isolated by Phage Display

Viruses

Dawei Zhang¹  Yuan Mao²  Qing Cao¹  Lin Xiong³  Juan Wen¹  Renjie Chen¹ 

[1]Department of Otolaryngology Head and Neck Surgery, The Second Affiliated Hospital of Nanjing Medical University, 121 Jiangjiayuan, Nanjing 210011, China

[2] E-Mails:

[3]Department of Otolaryngology Head and Neck Surgery, Jiangsu Province Official Hospital, 65 Jiangsu Road, 210029 Nanjing, China

[4] E-Mail:

[5]Department of Pathology, The Second Affiliated Hospital of Nanjing Medical University, 121 Jiangjiayuan, Nanjing 210011, China

[6] E-Mail:

关键词: EBV;    LMP1;    phage antibody library;    nasopharyngeal carcinoma;    Fab antibody;   

DOI  :  10.3390/v5041131

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2013 by the authors; licensee MDPI, Basel, Switzerland.