NDLI: 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

Content Provider	Springer Nature : BioMed Central
Author	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
Abstract	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.
Related Links	https://virologyj.biomedcentral.com/counter/pdf/10.1186/s12985-018-1104-6.pdf
Ending Page	12
Page Count	12
Starting Page	1
File Format	HTM / HTML
DOI	10.1186/s12985-018-1104-6
Journal	Virology Journal
Issue Number	1
Volume Number	15
Language	English
Publisher	BioMed Central
Publisher Date	2018-12-27
Access Restriction	Open
Subject Keyword	Virology Zika virus Envelope protein CD4 fusion tag Protein production ELISA Mexican patients Neutralizing antibodies
Content Type	Text
Resource Type	Article
Subject	Virology Infectious Diseases
Journal Impact Factor	4/2023
5-Year Journal Impact Factor	3.8/2023

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