Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2019 Sep;35(5):e2831.
doi: 10.1002/btpr.2831. Epub 2019 May 29.

Development of an alternating tangential flow (ATF) perfusion-based transient gene expression (TGE) bioprocess for universal influenza vaccine

Jinsung Hong  1 , Jacob Demirji  1 , Daniel Blackstock  1 , James Lee  1 , Tracey Dinh  1 , Alvenne Goh  1 , Frank Arnold  1 , Joe Horwitz  1
Affiliations

Development of an alternating tangential flow (ATF) perfusion-based transient gene expression (TGE) bioprocess for universal influenza vaccine

Jinsung Hong et al. Biotechnol Prog. 2019 Sep.

Abstract

An alternating tangential flow (ATF) perfusion-based transient gene expression (TGE) bioprocess has been developed using human embryonic kidney (HEK) 293 cells to produce H1-ss-np, a promising candidate for a universal influenza vaccine. Two major adjustments were taken to improve the process: (1) eliminate the interference of microbubbles during gene transfection; and (2) utilize an ATF perfusion system for a prolonged culture period. As a result, a closed-operation 9-days ATF perfusion-based TGE bioprocess was developed. The TGE bioprocess showed continuous cell growth with high cell viability and prolonged cellular productivity that achieved recombinant product level of ~270 mg/L which was more than two times that of 4-days base-line TGE bioprocess. In addition, the consumables cost per milligram for ATF perfusion-based TGE bioprocess was ~70% lower than that of the base-line TGE bioprocess suggesting high cost savings potential in vaccine manufacturing. Based on the lower contamination risk, higher productivity, and cost efficiency, the ATF perfusion-based TGE bioprocess can likely provide potential benefits to many future applications in vaccine and drug manufacturing.

Keywords: alternating tangential flow (ATF); mammalian cell culture; perfusion bioreactor; transient gene expression (TGE).

PubMed Disclaimer

References

REFERENCES

    1. Wright JF. Transient transfection methods for clinical adeno-associated viral vector production. Hum Gene Ther. 2009;20(7):698-706.
    1. Khan KH. Gene expression in mammalian cells and its applications. Adv Pharm Bull. 2013;3(2):257-263.
    1. Girard P, Derouazi M, Baumgartner G, et al. 100-liter transient transfection. Cytotechnology. 2002;38(1-3):15-21.
    1. Grieger JC, Soltys SM, Samulski RJ. Production of recombinant Adeno-associated virus vectors using suspension HEK293 cells and continuous harvest of vector from the culture media for GMP FIX and FLT1 clinical vector. Mol Ther. 2016;24(2):287-297.
    1. Baldi L, Hacker DL, Adam M, Wurm FM. Recombinant protein production by large-scale transient gene expression in mammalian cells: state of the art and future perspectives. Biotechnol Lett. 2007;29(5):677-684.

LinkOut - more resources