title: Generation of monoclonal antibodies against native viral proteins using antigen-expressing mammalian cells for mouse immunization.
abstract: Due 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.
  
  
  
  Here 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.
  
  
  
  The 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.
authors.affiliation: Swiss Tropical and Public Health Institute, Molecular Immunology, Basel, Switzerland.|University of Basel, Basel, Switzerland.
authors.forename: Natalie
authors.initials: N
authors.lastname: Rose
authors.affiliation: Biology Institute, Fluminense Federal University, Laboratory of Molecular Virology, Niterói, Brazil.
authors.forename: Carlos Augusto
authors.initials: CA
authors.lastname: Pinho-Nascimento
authors.affiliation: Department of Infectious Diseases, University of Heidelberg, Molecular Virology, Heidelberg, Germany.
authors.forename: Alessia
authors.initials: A
authors.lastname: Ruggieri
authors.affiliation: Swiss Tropical and Public Health Institute, Molecular Immunology, Basel, Switzerland.|University of Basel, Basel, Switzerland.
authors.forename: Paola
authors.initials: P
authors.lastname: Favuzza
authors.affiliation: Swiss Tropical and Public Health Institute, Molecular Immunology, Basel, Switzerland.|University of Basel, Basel, Switzerland.
authors.forename: Marco
authors.initials: M
authors.lastname: Tamborrini
authors.affiliation: Department of Infectious Diseases, University of Heidelberg, Molecular Virology, Heidelberg, Germany.
authors.forename: Hanna
authors.initials: H
authors.lastname: Roth
authors.affiliation: Oswaldo Cruz Institute, Fiocruz, Laboratory of comparative and environmental Virology, Rio de Janeiro, Brazil.
authors.forename: Marcia Terezinha
authors.initials: MT
authors.lastname: Baroni de Moraes
authors.affiliation: Swiss Tropical and Public Health Institute, Molecular Immunology, Basel, Switzerland.|University of Basel, Basel, Switzerland.
authors.forename: Hugues
authors.initials: H
authors.lastname: Matile
authors.affiliation: Section Clinical Tropical Medicine, Department of Infectious Diseases, Heidelberg University Hospital, Heidelberg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Jänisch
authors.affiliation: Swiss Tropical and Public Health Institute, Molecular Immunology, Basel, Switzerland.|University of Basel, Basel, Switzerland.
authors.forename: Gerd
authors.initials: G
authors.lastname: Pluschke
authors.affiliation: Swiss Tropical and Public Health Institute, Molecular Immunology, Basel, Switzerland. Katharina.roeltgen@unibas.ch.|University of Basel, Basel, Switzerland. Katharina.roeltgen@unibas.ch.
authors.forename: Katharina
authors.initials: K
authors.lastname: Röltgen
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000938:Antigen-Presenting Cells / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology*; D015195:Drug Design; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D018604:Epitope Mapping; D057809:HEK293 Cells; D006801:Humans; D007114:Immunization / Q000379:methods*; D007118:Immunoassay / Q000379:methods; D051379:Mice; D017361:Viral Nonstructural Proteins / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; C082515:NS1 protein, dengue-1 virus; D017361:Viral Nonstructural Proteins
keywords: Dengue virus; HEK cells; Hybridoma technology; Monoclonal antibodies; Mouse immunization; NS1 protein; Transfection