Production and Purification of Zika Virus NS1 Glycoprotein in HEK293 Cells

Methods Mol Biol. 2020:2142:93-102. doi: 10.1007/978-1-0716-0581-3_8.

Abstract

In this chapter, we describe production and purification of the Zika virus NS1 glycoprotein in human embryonic kidney (HEK293T) cells at small, research laboratory scale. The expression of secreted NS1 (sNS1) and the C-terminal β-ladder domain in HEK293T cells were tested in a small-scale transfection before scaling up to a larger-scale transfection using roller bottles. Two different purification approaches have been applied to obtain purified NS1 (sNS1) and the C-terminal β-ladder domain ready for clinical applications.

Keywords: HEK293T; NS1 glycoprotein; Protein production; Protein purification; Zika virus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques / methods
  • Cloning, Molecular / methods*
  • HEK293 Cells
  • Histidine / chemistry
  • Humans
  • Oligopeptides / chemistry
  • Proteomics / methods
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Secretory Pathway
  • Serologic Tests / methods
  • Transfection / methods*
  • Viral Nonstructural Proteins / genetics*
  • Viral Nonstructural Proteins / isolation & purification*
  • Viral Nonstructural Proteins / metabolism*
  • Zika Virus / genetics

Substances

  • His-His-His-His-His-His
  • NS1 protein, zika virus
  • Oligopeptides
  • Recombinant Proteins
  • Viral Nonstructural Proteins
  • Histidine