» Articles » PMID: 24713807

Enhanced Production of Chikungunya Virus-like Particles Using a High-pH Adapted Spodoptera Frugiperda Insect Cell Line

Overview
Journal PLoS One
Date 2014 Apr 10
PMID 24713807
Citations 19
Authors
Affiliations
Soon will be listed here.
Abstract

Chikungunya virus-like particles (VLPs) have potential to be used as a prophylactic vaccine based on testing in multiple animal models and are currently being evaluated for human use in a Phase I clinical trial. The current method for producing these enveloped alphavirus VLPs by transient gene expression in mammalian cells presents challenges for scalable and robust industrial manufacturing, so the insect cell baculovirus expression vector system was evaluated as an alternative expression technology. Subsequent to recombinant baculovirus infection of Sf21 cells in standard culture media (pH 6.2-6.4), properly processed Chikungunya structural proteins were detected and assembled capsids were observed. However, an increase in culture pH to 6.6-6.8 was necessary to produce detectable concentrations of assembled VLPs. Since this elevated production pH exceeds the optimum for growth medium stability and Sf21 culture, medium modifications were made and a novel insect cell variant (SfBasic) was derived by exposure of Sf21 to elevated culture pH for a prolonged period of time. The high-pH adapted SfBasic insect cell line described herein is capable of maintaining normal cell growth into the typical mammalian cell culture pH range of 7.0-7.2 and produces 11-fold higher Chikungunya VLP yields relative to the parental Sf21 cell line. After scale-up into stirred tank bioreactors, SfBasic derived VLPs were chromatographically purified and shown to be similar in size and structure to a VLP standard derived from transient gene expression in HEK293 cells. Total serum anti-Chikungunya IgG and neutralizing titers from guinea pigs vaccinated with SfBasic derived VLPs or HEK293 derived VLPs were not significantly different with respect to production method, suggesting that this adapted insect cell line and production process could be useful for manufacturing Chikungunya VLPs for use as a vaccine. The adaptation of Sf21 to produce high levels of recombinant protein and VLPs in an elevated pH range may also have applications for other pH-sensitive protein or VLP targets.

Citing Articles

Serum-Free Suspension Culture of the C6/36 Cell Line for Chimeric Orthoflavivirus Vaccine Production.

Dawurung J, Harrison J, Modhiran N, Hall R, Hobson-Peters J, de Malmanche H Viruses. 2025; 17(2).

PMID: 40007005 PMC: 11860912. DOI: 10.3390/v17020250.


Adaptive Laboratory Evolution to Improve Recombinant Protein Production Using Insect Cells.

Correia R, Fernandes B, Alves P, Roldao A Methods Mol Biol. 2024; 2829:79-90.

PMID: 38951328 DOI: 10.1007/978-1-0716-3961-0_6.


Pathogenesis of Rift Valley Fever Virus in a BALB/c Mouse Model Is Affected by Virus Culture Conditions and Sex of the Animals.

Graham V, Easterbrook L, Kennedy E, Rayner E, Findlay-Wilson S, Flett L Viruses. 2023; 15(12).

PMID: 38140610 PMC: 10747589. DOI: 10.3390/v15122369.


Zika virus-like particles (VLPs) produced in insect cells.

de Mello R, Consoni Bernardino T, Oliveira Guardalini L, Mancini Astray R, Antoniazzi M, Jared S Front Pharmacol. 2023; 14:1181566.

PMID: 37377933 PMC: 10291072. DOI: 10.3389/fphar.2023.1181566.


Size-selective downstream processing of virus particles and non-enveloped virus-like particles.

Hillebrandt N, Hubbuch J Front Bioeng Biotechnol. 2023; 11:1192050.

PMID: 37304136 PMC: 10248422. DOI: 10.3389/fbioe.2023.1192050.


References
1.
Gerbal M, Fournier P, Barry P, Mariller M, Odier F, Devauchelle G . Adaptation of an insect cell line of Spodoptera frugiperda to grow at 37 degrees C: characterization of an endodiploid clone. In Vitro Cell Dev Biol Anim. 2000; 36(2):117-24. DOI: 10.1290/1071-2690(2000)036<0117:aoaicl>2.0.co;2. View

2.
Sorzano C, Marabini R, Velazquez-Muriel J, Bilbao-Castro J, Scheres S, Carazo J . XMIPP: a new generation of an open-source image processing package for electron microscopy. J Struct Biol. 2004; 148(2):194-204. DOI: 10.1016/j.jsb.2004.06.006. View

3.
Lander G, Stagg S, Voss N, Cheng A, Fellmann D, Pulokas J . Appion: an integrated, database-driven pipeline to facilitate EM image processing. J Struct Biol. 2009; 166(1):95-102. PMC: 2775544. DOI: 10.1016/j.jsb.2009.01.002. View

4.
Marquardt M, Phalen T, Kielian M . Cholesterol is required in the exit pathway of Semliki Forest virus. J Cell Biol. 1993; 123(1):57-65. PMC: 2119816. DOI: 10.1083/jcb.123.1.57. View

5.
Lipin D, Chuan Y, Lua L, Middelberg A . Encapsulation of DNA and non-viral protein changes the structure of murine polyomavirus virus-like particles. Arch Virol. 2008; 153(11):2027-39. DOI: 10.1007/s00705-008-0220-9. View