Journal Article

Cloning and expression of two human recombinant monoclonal Fab fragments specific for EBV viral capsid antigen

Lingli Dong, Yasufumi Masaki, Tsutomu Takegami, Takafumi Kawanami, Kunihiko Itoh, Zhe-Xiong Jin, Cheng-Ri Huang, Xiao-Peng Tong, Toshihiro Fukushima, Masao Tanaka, Toshioki Sawaki, Tomoyuki Sakai, Susumu Sugai, Toshiro Okazaki, Yuko Hirose and Hisanori Umehara

in International Immunology

Published on behalf of Japanese Society for Immunology

Volume 19, issue 3, pages 331-336
Published in print March 2007 | ISSN: 0953-8178
Published online January 2007 | e-ISSN: 1460-2377 | DOI: http://dx.doi.org/10.1093/intimm/dxl150
Cloning and expression of two human recombinant monoclonal Fab fragments specific for EBV viral capsid antigen

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Serum titers of antibody to Epstein–Barr virus (EBV) viral capsid antigen (VCA) have been positively correlated with malignancies of lymphoid proliferation, such as Burkitt's lymphoma and Hodgkin's lymphoma. We have constructed a phage display combinatorial antibody Fab library from a patient with marginal zone B cell lymphoma associated with Sjögren's syndrome and carrying high serum anti-EBV-VCA IgG titer. Fab fragments were selected by panning against EBV-VCA protein coated onto ELISA plates, and selected Fab clones were characterized by ELISA, western blotting (WB), indirect immunofluorescence assay and immunohistochemistry. We have established two Fab clones, Fab-aVCA1 and Fab-aVCA21, which specifically recognize EBV-VCA by ELISA and WB. Inhibition ELISA competition showed that both clones could significantly reduce the binding of specific anti-EBV-VCA mAb to its relevant proteins. Furthermore, these two Fab clones could localize VCA protein in the EBV-positive P3HR1 and Daudi cell lines, as well as in tissue samples from patients with EBV-infected lymphoid malignancies. These results indicate that our two Fab clones are novel human mAbs specific for EBV-VCA protein and may have potential benefits for development of novel diagnostic and therapeutic approaches in EBV-related lymphoid malignancies.

Keywords: antibodies; human; molecular biology

Journal Article.  3395 words.  Illustrated.

Subjects: Immunology ; Biological Sciences

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