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

نویسندگان

  • 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
  • Hisanori Umehara
چکیده

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.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Production of monoclonal antibody against recombinant NS3 protein of bovine viral diarrhea virus (NADL strain)

Bovine Viral Diarrhea virus (BVDV) is an important viral pathogen of cattle causing several clinical syndromes. There are usually no pathognomonic clinical signs of BVDV infection. Diagnostic investigations therefore rely on serological detection and virus isolation. Nonstructural protein 3 (NS3) as immunogenic protein of BVDV is genetically and antigenically conserved among different isolates....

متن کامل

Cloning and Expression of Coxsakievirus B3 Viral Protein-1 in E. Coli

Viral protein-1 (VP1) is a major capsid protein of Coxsakievirus B3 (CVB3) that plays an important role in directing viruses towards permissive cells and acts as a main antigenic site of the virus in eliciting of host immune response, hence it seems VP1 can be considered as a vaccine candidate against CVB3 infection. In this study, cDNA of VP1 was prepared, cloned into pET expression vector and...

متن کامل

Construction and cloning of a recombinant expression vector containing human Cd20 Gene for antibody therapy in Non-Hodgkin Lymphoma

ABSTRACT Introduction: Non-Hodgkin lymphoma (NHL) is a cancer that starts in lymphocytes. The main treatment for NHL is chemotherapy and radiation. Today immunotherapy is a promising therapeutic approach in the treatment of a variety cancers which is high specific unlike previous methods. Antibodies do not penetrate effectively into tumore tissues because of their large size. Whe...

متن کامل

Cloning and Expression of Recombinant Camelid Single-Domain Antibody in Tobacco

Antibodies provide a suitable tool in fundamental research and their high affinity and specificity make them invaluable for diagnostic and therapeutic applications. A promising alternative to conventional antibodies are the heavy chain antibodies (VHH) of Camelidae having short length, high solubility and stability are preferred to other antibody derivatives. In this study, our goal was product...

متن کامل

Human Papillomavirus Type16- L1 VLP Production in Insect Cells

  Objective(s):  Infection by high-risk papillomavirus is regarded as the major risk factor in the development of cervical cancer. Recombinant DNA technology allows expression of the L1 major capsid protein of HPV in different expression systems, which has intrinsic capacity to self-assemble into viral-like particles (VLP). VLPS are non-infectious, highly immunogenic and can elicit neutralizing...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • International immunology

دوره 19 3  شماره 

صفحات  -

تاریخ انتشار 2007