Purification of Recombinant Ebola Virus Glycoprotein and VP40 from a Human Cell Line
Technical Report,01 Apr 2015,31 Jan 2016
Excet, Inc. Gunpowder United States
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The recent, widespread Ebola virus outbreak that originated in West Africa underscores the need for novel diagnostic and therapeutic tools to treat and contain future outbreaks. In line with this goal, the in-house production of Ebola antigens allows researchers to develop and test novel reagents without the requirement for high-level biological containment. In this report, we describe procedures for the recombinant expression and purification of the Ebola Zaire structural glycoprotein and matrix protein VP40 from a human cell line. Plasmids coding for the expression of these proteins were transiently transfected into human embryonic kidney cells 293 and secreted into the culture medium. Next, purification proceeded using immobilized metal affinity chromatography and anion exchange chromatography. The typical yield for each of these proteins was approximately 1 mg30 mL of cell culture with 90 purity. The procedures described herein may be useful for research efforts involving antibody discovery and immunoassay development targeting the Ebola virus.
- Genetic Engineering and Molecular Biology
- Medicine and Medical Research