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Zika virus (ZIKV) is an emerging mosquito-borne flavivirus, which has recently caused global epidemics with its association with congenital Zika syndrome such as severe microcephaly. The recombinant ZIKV envelope (Env) glycoprotein is useful for immunological applications such as serodiagnosis of ZIKV infection and for monitoring immune responses in preclinical and clinical ZIKV vaccine developments. In this chapter, we describe the optimization of production of Zika virus envelope glycoprotein in Human Embryonic Kidney (HEK 293T) cells by small-scale expression followed by large-scale protein production. Small-scale expression of HEK 293T cells allows screening of a large number of vectors simultaneously to select the vectors with best secretory profiles for scale-up in Expi293 mammalian system to maximize the protein yield followed by purification for research and clinical applications.

Original publication

DOI

10.1007/978-1-0716-0581-3_9

Type

Journal article

Journal

Methods in molecular biology (Clifton, N.J.)

Publication Date

01/2020

Volume

2142

Pages

103 - 112

Addresses

The Jenner Institute, The Henry Wellcome Building for Molecular Physiology, Nuffield Department of Medicine, University of Oxford, Oxford, UK.

Keywords

Humans, Gene Products, env, Recombinant Fusion Proteins, Viral Envelope Proteins, Enzyme-Linked Immunosorbent Assay, Serologic Tests, Calibration, Transfection, Proteomics, Gene Expression, Secretory Pathway, HEK293 Cells, Zika Virus, CD4 Antigens, Viral Envelope