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Optimization of Zika virus envelope protein production for ELISA and correlation of antibody titers with virus neutralization in Mexican patients from an arbovirus endemic region.
Kim, Young Chan; Lopez-Camacho, Cesar; Nettleship, Joanne E; Rahman, Nahid; Hill, Michelle L; Silva-Reyes, Laura; Ortiz-Martinez, Georgina; Figueroa-Aguilar, Gloria; Mar, María Antonieta; Vivanco-Cid, Héctor; Rollier, Christine S; Zitzmann, Nicole; Viveros-Sandoval, Martha Eva; Owens, Raymond J; Reyes-Sandoval, Arturo.
Afiliação
  • Kim YC; The Jenner Institute, Nuffield Department of Medicine, The Henry Wellcome Building for Molecular Physiology, University of Oxford, Old Road Campus Research Building. Roosevelt Drive, Oxford, OX3 7DQ, UK.
  • Lopez-Camacho C; Division of Structural Biology, Wellcome Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, UK.
  • Nettleship JE; The Jenner Institute, Nuffield Department of Medicine, The Henry Wellcome Building for Molecular Physiology, University of Oxford, Old Road Campus Research Building. Roosevelt Drive, Oxford, OX3 7DQ, UK.
  • Rahman N; Division of Structural Biology, Wellcome Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, UK.
  • Hill ML; Rutherford Appleton Laboratory, OPPF-UK, Research Complex at Harwell, Oxford, UK.
  • Silva-Reyes L; Division of Structural Biology, Wellcome Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, UK.
  • Ortiz-Martinez G; Rutherford Appleton Laboratory, OPPF-UK, Research Complex at Harwell, Oxford, UK.
  • Figueroa-Aguilar G; Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
  • Mar MA; Oxford Vaccine Group, Department of Paediatrics, University of Oxford and the NIHR Oxford Biomedical Research Centre, Centre for Clinical Vaccinology and Tropical Medicine, Churchill Hospital, Oxford, UK.
  • Vivanco-Cid H; Laboratorio de Hemostasia y Biología Vascular. División de Estudios de Posgrado. Facultad de Ciencias Médicas y Biológicas "Dr. Ignacio Chávez", Universidad Michoacana de San Nicolás de Hidalgo, UMSNH, Morelia, Mexico.
  • Rollier CS; UMSNH-Oxford University of Oxford Clinical Research Laboratory (UMOCRL), Faculty of Biological and Medical Sciences "Dr. Ignacio Chávez", Universidad Michoacana de San Nicolás de Hidalgo, Morelia, Michoacán, Mexico.
  • Zitzmann N; Laboratorio Estatal de Salud Pública, Secretaría de Salud de Michoacán, Morelia, Michoacán, Mexico.
  • Viveros-Sandoval ME; HGZMF No. 12 Lázaro Cárdenas Michoacán dirección av. Lázaro Cárdenas No. 154 Col. Centro Lázaro Cárdenas Michoacán, Veracruz, Mexico.
  • Owens RJ; Instituto de Investigaciones Médico-Biológicas, Universidad Veracruzana, Veracruz, Mexico.
  • Reyes-Sandoval A; Oxford Vaccine Group, Department of Paediatrics, University of Oxford and the NIHR Oxford Biomedical Research Centre, Centre for Clinical Vaccinology and Tropical Medicine, Churchill Hospital, Oxford, UK.
Virol J ; 15(1): 193, 2018 12 27.
Article em En | MEDLINE | ID: mdl-30587198
BACKGROUND: Zika virus (ZIKV) has become a global threat with immediate need for accurate diagnostics, efficacious vaccines and therapeutics. Several ZIKV envelope (Env)-based vaccines have been developed recently. However, many commercially available ZIKV Env are based on the African lineage and produced in insect cells. Here, we sought to produce Asian-lineage ZIKV Env in mammalian cells for research and clinical applications. METHODS: We designed various gene expression constructs to optimize the production of ZIKV using prM-Env and full or C-terminal truncations of Env; with or without a rat CD4 fusion partner to allow large-scale production of soluble protein in mammalian HEK293 cells. Protein expression was verified by mass spectrometry and western-blot with a pan-flavivirus antibody, a ZIKV Env monoclonal antibody and with immune sera from adenoviral (ChAdOx1) ZIKV Env-vaccinated mice. The resulting Env-CD4 was used as a coating reagent for immunoassay (ELISA) using both mouse and human seropositive sera. RESULTS: Replacement of the C-terminus transmembrane Env domain by a rat CD4 and addition of prM supported optimal expression and secretion of Env. Binding between the antigens and the antibodies was similar to binding when using commercially available ZIKV Env reagents. Furthermore, antibodies from ZIKV patients bound ZIKV Env-CD4 in ELISA assays, whereas sera from healthy blood donors yielded minimal OD background. The serological outcomes of this assay correlated also with ZIKV neutralisation capacity in vitro. CONCLUSIONS: Results obtained from this study indicate the potential of the Asian-lineage Zika Env-CD4 and Env proteins in ELISA assays to monitor humoral immune responses in upcoming clinical trials as well as a sero-diagnostic tool in ZIKV infection.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas Recombinantes de Fusão / Proteínas do Envelope Viral / Anticorpos Neutralizantes / Zika virus / Anticorpos Antivirais Limite: Animals / Humans País/Região como assunto: Mexico Idioma: En Revista: Virol J Assunto da revista: VIROLOGIA Ano de publicação: 2018 Tipo de documento: Article País de publicação: Reino Unido
Texto completo
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XML
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas Recombinantes de Fusão / Proteínas do Envelope Viral / Anticorpos Neutralizantes / Zika virus / Anticorpos Antivirais Limite: Animals / Humans País/Região como assunto: Mexico Idioma: En Revista: Virol J Assunto da revista: VIROLOGIA Ano de publicação: 2018 Tipo de documento: Article País de publicação: Reino Unido