Generation of monoclonal antibodies against native viral proteins using antigen-expressing mammalian cells for mouse immunization

BMC Biotechnology

Methodology Article

Carlos Augusto Pinho-Nascimento¹  Alessia Ruggieri²  Hanna Roth²  Marcia Terezinha Baroni de Moraes³  Thomas Jänisch⁴  Hugues Matile⁵  Gerd Pluschke⁵  Paola Favuzza⁵  Marco Tamborrini⁵  Katharina Röltgen⁵  Natalie Rose⁵ 

[1] Biology Institute, Fluminense Federal University, Laboratory of Molecular Virology, Niterói, Brazil;Department of Infectious Diseases, University of Heidelberg, Molecular Virology, Heidelberg, Germany;Oswaldo Cruz Institute, Fiocruz, Laboratory of comparative and environmental Virology, Rio de Janeiro, Brazil;Section Clinical Tropical Medicine, Department of Infectious Diseases, Heidelberg University Hospital, Heidelberg, Germany;Swiss Tropical and Public Health Institute, Molecular Immunology, Basel, Switzerland;University of Basel, Basel, Switzerland;

关键词: Dengue virus;    NS1 protein;    Mouse immunization;    HEK cells;    Transfection;    Hybridoma technology;    Monoclonal antibodies;   

DOI  :  10.1186/s12896-016-0314-5

 received in 2016-08-01, accepted in 2016-11-01,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundDue to their rising incidence and progressive geographical spread, infections with mosquito-borne viruses, such as dengue (DENV), chikungunya and zika virus, have developed into major public health challenges. Since all of these viruses may cause similar symptoms and can occur in concurrent epidemics, tools for their differential diagnosis and epidemiological monitoring are of urgent need.ResultsHere we report the application of a novel strategy to rapidly generate monoclonal antibodies (mAbs) against native viral antigens, exemplified for the DENV nonstructural glycoprotein 1 (NS1). The described system is based on the immunization of mice with transfected mammalian cells expressing the target antigens in multiple displays on their cell surface and thereby presenting them efficiently to the host immune system in their native conformation. By applying this cell-based approach to the DENV NS1 protein of serotypes 1 (D1NS1) and 4 (D4NS1), we were able to rapidly generate panels of DENV NS1 serotype cross-reactive, as well as D1NS1- and D4NS1 serotype-specific mAbs. Our data show that the generated mAbs were capable of recognizing the endogenous NS1 protein in DENV-containing biological samples.ConclusionThe use of this novel immunization strategy, allows for a fast and efficient generation of hybridoma cell lines, producing mAbs against native viral antigens. Envisaged applications of the mAbs include the development of test platforms enabling a differentiation of the DENV serotypes and high resolution immunotyping for epidemiological studies.

【 授权许可】

CC BY   
© The Author(s). 2016