Publication: Generation and characterization of cross neutralizing human monoclonal antibody against 4 serotypes of dengue virus without enhancing activity
Issued Date
2017-01-01
Resource Type
ISSN
21678359
Other identifier(s)
2-s2.0-85033579945
Rights
Mahidol University
Rights Holder(s)
SCOPUS
Bibliographic Citation
PeerJ. Vol.2017, No.11 (2017)
Suggested Citation
Subenya Injampa, Nataya Muenngern, Chonlatip Pipattanaboon, Surachet Benjathummarak, Khwanchit Boonha, Hathairad Hananantachai, Waranya Wongwit, Pongrama Ramasoota, Pannamthip Pitaksajjakul Generation and characterization of cross neutralizing human monoclonal antibody against 4 serotypes of dengue virus without enhancing activity. PeerJ. Vol.2017, No.11 (2017). doi:10.7717/peerj.4021 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/41538
Research Projects
Organizational Units
Authors
Journal Issue
Thesis
Title
Generation and characterization of cross neutralizing human monoclonal antibody against 4 serotypes of dengue virus without enhancing activity
Other Contributor(s)
Abstract
© 2017 Injampa et al. Background. Dengue disease is a leading cause of illness and death in the tropics and subtropics. Most severe cases occur among patients secondarily infected with a different dengue virus (DENV) serotype compared with that from the first infection, resulting in antibody-dependent enhancement activity (ADE). Our previous study generated the neutralizing human monoclonal antibody, D23-1B3B9 (B3B9), targeting the first domain II of E protein, which showed strong neutralizing activity (NT) against all four DENV serotypes. However, at sub-neutralizing concentrations, it showed ADE activity in vitro. Methods. In this study, we constructed a new expression plasmid using the existing IgG heavy chain plasmid as a template for Fc modification at position N297Q bysite-directed mutagenesis. The resulting plasmid was then co-transfected with a light chain plasmid to produce full recombinant IgG (rIgG) in mammalian cells(N297Q-B3B9). This rIgG was characterized for neutralizing and enhancing activity by using different Fc R bearing cells. To produce sufficient quantities of B3B9 rIgG for further characterization, CHO-K1 cells stably secreting N297Q-B3B9 rIgG were then established. Results. The generated N297Q-B3B9 rIgG which targets the conserved N-terminal fusion loop ofDENVenvelope protein showed the same cross-neutralizing activity to all four DENV serotypes as those of wild type rIgG. In both FcγRI- and RII-bearing THP- 1 cells and FcγRII-bearing K562 cells, N297Q-B3B9 rIgG lacked ADE activity against all DENV serotypes at sub-neutralizing concentrations. Fortunately, the N297Q-B3B9rIgG secreted from stable cells showed the same patterns of NT and ADE activities as those of the N297Q-B3B9 rIgG obtained from transient expression against DENV2. Thus, the CHO-K1 stably expressing N297Q-B3B9 HuMAb can be developed as high producer stable cells and used to produce sufficient amounts of antibody for further characterization as a promising dengue therapeutic candidate. Discussion. Human monoclonal antibody, targeted to fusion loop of envelope domainII (EDII), was generated and showed cross-neutralizing activity to 4 serotypes of DENV, but did not cause any viral enhancement activity in vitro. This HuMAb could be further developed as therapeutic candidates. Subjects Biotechnology, Molecular Biology, Virology, Infectious Diseases.