Efficient production of foot-and-mouth disease virus empty capsids in insect cells following down regulation of 3C protease activity

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Efficient production of foot-and-mouth disease virus empty capsids in insect cells following down regulation of 3C protease activity. / Porta, Claudine; Xu, Xiaodong; Loureiro, Silvia; Paramasivam, Saravanan; Ren, Junyuan; Al-Khalil, Tara; Burman, Alison; Jackson, Terry; Belsham, Graham J; Curry, Stephen; Lomonossoff, George P; Parida, Satya; Paton, David; Li, Yanmin; Wilsden, Ginette; Ferris, Nigel; Owens, Ray; Kotecha, Abhay; Fry, Elizabeth; Stuart, David I; Charleston, Bryan; Jones, Ian M.

In: Journal of Virological Methods, Vol. 187, No. 2, 02.2013, p. 406-12.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Porta, C, Xu, X, Loureiro, S, Paramasivam, S, Ren, J, Al-Khalil, T, Burman, A, Jackson, T, Belsham, GJ, Curry, S, Lomonossoff, GP, Parida, S, Paton, D, Li, Y, Wilsden, G, Ferris, N, Owens, R, Kotecha, A, Fry, E, Stuart, DI, Charleston, B & Jones, IM 2013, 'Efficient production of foot-and-mouth disease virus empty capsids in insect cells following down regulation of 3C protease activity', Journal of Virological Methods, vol. 187, no. 2, pp. 406-12. https://doi.org/10.1016/j.jviromet.2012.11.011

APA

Porta, C., Xu, X., Loureiro, S., Paramasivam, S., Ren, J., Al-Khalil, T., Burman, A., Jackson, T., Belsham, G. J., Curry, S., Lomonossoff, G. P., Parida, S., Paton, D., Li, Y., Wilsden, G., Ferris, N., Owens, R., Kotecha, A., Fry, E., ... Jones, I. M. (2013). Efficient production of foot-and-mouth disease virus empty capsids in insect cells following down regulation of 3C protease activity. Journal of Virological Methods, 187(2), 406-12. https://doi.org/10.1016/j.jviromet.2012.11.011

Vancouver

Porta C, Xu X, Loureiro S, Paramasivam S, Ren J, Al-Khalil T et al. Efficient production of foot-and-mouth disease virus empty capsids in insect cells following down regulation of 3C protease activity. Journal of Virological Methods. 2013 Feb;187(2):406-12. https://doi.org/10.1016/j.jviromet.2012.11.011

Author

Porta, Claudine ; Xu, Xiaodong ; Loureiro, Silvia ; Paramasivam, Saravanan ; Ren, Junyuan ; Al-Khalil, Tara ; Burman, Alison ; Jackson, Terry ; Belsham, Graham J ; Curry, Stephen ; Lomonossoff, George P ; Parida, Satya ; Paton, David ; Li, Yanmin ; Wilsden, Ginette ; Ferris, Nigel ; Owens, Ray ; Kotecha, Abhay ; Fry, Elizabeth ; Stuart, David I ; Charleston, Bryan ; Jones, Ian M. / Efficient production of foot-and-mouth disease virus empty capsids in insect cells following down regulation of 3C protease activity. In: Journal of Virological Methods. 2013 ; Vol. 187, No. 2. pp. 406-12.

Bibtex

@article{8af7c64181b040c497e671e3800d3926,

title = "Efficient production of foot-and-mouth disease virus empty capsids in insect cells following down regulation of 3C protease activity",

abstract = "Foot-and-mouth disease virus (FMDV) is a significant economically and distributed globally pathogen of Artiodactyla. Current vaccines are chemically inactivated whole virus particles that require large-scale virus growth in strict bio-containment with the associated risks of accidental release or incomplete inactivation. Non-infectious empty capsids are structural mimics of authentic particles with no associated risk and constitute an alternate vaccine candidate. Capsids self-assemble from the processed virus structural proteins, VP0, VP3 and VP1, which are released from the structural protein precursor P1-2A by the action of the virus-encoded 3C protease. To date recombinant empty capsid assembly has been limited by poor expression levels, restricting the development of empty capsids as a viable vaccine. Here expression of the FMDV structural protein precursor P1-2A in insect cells is shown to be efficient but linkage of the cognate 3C protease to the C-terminus reduces expression significantly. Inactivation of the 3C enzyme in a P1-2A-3C cassette allows expression and intermediate levels of 3C activity resulted in efficient processing of the P1-2A precursor into the structural proteins which assembled into empty capsids. Expression was independent of the insect host cell background and leads to capsids that are recognised as authentic by a range of anti-FMDV bovine sera suggesting their feasibility as an alternate vaccine.",

keywords = "Animals, Biotechnology/methods, Capsid/immunology, Cell Line, Cysteine Endopeptidases/biosynthesis, Down-Regulation, Foot-and-Mouth Disease Virus/genetics, Gene Expression, Insecta, Technology, Pharmaceutical/methods, Viral Proteins/biosynthesis, Viral Vaccines/genetics",

author = "Claudine Porta and Xiaodong Xu and Silvia Loureiro and Saravanan Paramasivam and Junyuan Ren and Tara Al-Khalil and Alison Burman and Terry Jackson and Belsham, {Graham J} and Stephen Curry and Lomonossoff, {George P} and Satya Parida and David Paton and Yanmin Li and Ginette Wilsden and Nigel Ferris and Ray Owens and Abhay Kotecha and Elizabeth Fry and Stuart, {David I} and Bryan Charleston and Jones, {Ian M}",

year = "2013",

month = feb,

doi = "10.1016/j.jviromet.2012.11.011",

language = "English",

volume = "187",

pages = "406--12",

journal = "Journal of Virological Methods",

issn = "0166-0934",

publisher = "Elsevier",

number = "2",

}

RIS

TY - JOUR

T1 - Efficient production of foot-and-mouth disease virus empty capsids in insect cells following down regulation of 3C protease activity

AU - Porta, Claudine

AU - Xu, Xiaodong

AU - Loureiro, Silvia

AU - Paramasivam, Saravanan

AU - Ren, Junyuan

AU - Al-Khalil, Tara

AU - Burman, Alison

AU - Jackson, Terry

AU - Belsham, Graham J

AU - Curry, Stephen

AU - Lomonossoff, George P

AU - Parida, Satya

AU - Paton, David

AU - Li, Yanmin

AU - Wilsden, Ginette

AU - Ferris, Nigel

AU - Owens, Ray

AU - Kotecha, Abhay

AU - Fry, Elizabeth

AU - Stuart, David I

AU - Charleston, Bryan

AU - Jones, Ian M

PY - 2013/2

Y1 - 2013/2

N2 - Foot-and-mouth disease virus (FMDV) is a significant economically and distributed globally pathogen of Artiodactyla. Current vaccines are chemically inactivated whole virus particles that require large-scale virus growth in strict bio-containment with the associated risks of accidental release or incomplete inactivation. Non-infectious empty capsids are structural mimics of authentic particles with no associated risk and constitute an alternate vaccine candidate. Capsids self-assemble from the processed virus structural proteins, VP0, VP3 and VP1, which are released from the structural protein precursor P1-2A by the action of the virus-encoded 3C protease. To date recombinant empty capsid assembly has been limited by poor expression levels, restricting the development of empty capsids as a viable vaccine. Here expression of the FMDV structural protein precursor P1-2A in insect cells is shown to be efficient but linkage of the cognate 3C protease to the C-terminus reduces expression significantly. Inactivation of the 3C enzyme in a P1-2A-3C cassette allows expression and intermediate levels of 3C activity resulted in efficient processing of the P1-2A precursor into the structural proteins which assembled into empty capsids. Expression was independent of the insect host cell background and leads to capsids that are recognised as authentic by a range of anti-FMDV bovine sera suggesting their feasibility as an alternate vaccine.

AB - Foot-and-mouth disease virus (FMDV) is a significant economically and distributed globally pathogen of Artiodactyla. Current vaccines are chemically inactivated whole virus particles that require large-scale virus growth in strict bio-containment with the associated risks of accidental release or incomplete inactivation. Non-infectious empty capsids are structural mimics of authentic particles with no associated risk and constitute an alternate vaccine candidate. Capsids self-assemble from the processed virus structural proteins, VP0, VP3 and VP1, which are released from the structural protein precursor P1-2A by the action of the virus-encoded 3C protease. To date recombinant empty capsid assembly has been limited by poor expression levels, restricting the development of empty capsids as a viable vaccine. Here expression of the FMDV structural protein precursor P1-2A in insect cells is shown to be efficient but linkage of the cognate 3C protease to the C-terminus reduces expression significantly. Inactivation of the 3C enzyme in a P1-2A-3C cassette allows expression and intermediate levels of 3C activity resulted in efficient processing of the P1-2A precursor into the structural proteins which assembled into empty capsids. Expression was independent of the insect host cell background and leads to capsids that are recognised as authentic by a range of anti-FMDV bovine sera suggesting their feasibility as an alternate vaccine.

KW - Animals

KW - Biotechnology/methods

KW - Capsid/immunology

KW - Cell Line

KW - Cysteine Endopeptidases/biosynthesis

KW - Down-Regulation

KW - Foot-and-Mouth Disease Virus/genetics

KW - Gene Expression

KW - Insecta

KW - Technology, Pharmaceutical/methods

KW - Viral Proteins/biosynthesis

KW - Viral Vaccines/genetics

U2 - 10.1016/j.jviromet.2012.11.011

DO - 10.1016/j.jviromet.2012.11.011

M3 - Journal article

C2 - 23174161

VL - 187

SP - 406

EP - 412

JO - Journal of Virological Methods

JF - Journal of Virological Methods

SN - 0166-0934

IS - 2

ER -

ID: 257916853