Baculovirus expression and diagnostic utility of the glycoprotein E of bovine herpesvirus-1.1 Egyptian strain “Abu-Hammad”

El-Kholy, Alaa A.; Abdou, Eman R.; Rady, Douaa I.; Elseafy, Mai M.

doi:10.1016/j.jviromet.2013.03.024

Cited by 7 publications

(6 citation statements)

References 19 publications

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“…Baculovirus-mediated expression in insect cells has become well-established for the production of recombinant glycoproteins. Since several glycoproteins could readily be produced in many insect cell lines compared with mammalian cells [85], these insect cell lines were widely employed for glycoprotein expression [86][87][88]. Nevertheless, the conventional BES may not be the best tool for producing glycoproteins to generate complex VLPs especially for pharmaceutical purposes, as the insect cells-produced glycoproteins have clearly different N-glycans from those produced by mammalian cells [8,89,90].…”

Section: Glycosylation Of Recombinant Proteinsmentioning

confidence: 99%

Use of baculovirus expression system for generation of virus-like particles: Successes and challenges

Liu

et al. 2013

Protein Expression and Purification

106

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The baculovirus expression system (BES) has been one of the versatile platforms for the production of recombinant proteins requiring multiple post-translational modifications, such as folding, oligomerization, phosphorylation, glycosylation, acylation, disulfide bond formation and proteolytic cleavage. Advances in recombinant DNA technology have facilitated application of the BES, and made it possible to express multiple proteins simultaneously in a single infection and to produce multimeric proteins sharing functional similarity with their natural analogs. Therefore, the BES has been used for the production of recombinant proteins and the construction of virus-like particles (VLPs), as well as for the development of subunit vaccines, including VLP-based vaccines. The VLP, which consists of one or more structural proteins but no viral genome, resembles the authentic virion but cannot replicate in cells. The high-quality recombinant protein expression and post-translational modifications obtained with the BES, along with its capacity to produce multiple proteins, imply that it is ideally suited to VLP production. In this article, we critically review the pros and cons of using the BES as a platform to produce both enveloped and non-enveloped VLPs.

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Section: Glycosylation Of Recombinant Proteinsmentioning

confidence: 99%

Use of baculovirus expression system for generation of virus-like particles: Successes and challenges

Liu

et al. 2013

Protein Expression and Purification

106

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“…Reasons for using BacSF9 system for protein expression included (1) secretion into the growth medium; (2) production of less aggregated proteins with an increased probability of correct folding, and (3) post-translational modifications for better antigenicity and specificity. Previously, the baculovirus expression system was successfully used to express different viral proteins such as the tomato yellow leaf curl virus coat protein (TYLCV) (El-Gaied, Salem & Elmenofy, 2017), Garlic mite-borne filamentous virus (GarMbFV) (Ardisson-Arau'jo et al, 2013), and glycoprotein E (gE) of the Egyptian BoHV-1.1 Abu-Hammad strain (rBac/gE-AbuH) (El-Kholy et al, 2013;El-Gaied et al, 2020). Furthermore, recombinant proteins produced using this system were validated for their antigenicity.…”

Section: Discussionmentioning

confidence: 99%

Expression of 1B capsid protein of Foot-and-mouth disease virus (FMDV) using baculovirus expression system and its validation in detecting SAT 2- specific antisera

Elmenofy

Mohamed

El-Gaied

et al. 2020

PeerJ

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Foot-and-mouth disease virus (FMDV) is one of the most devastating animal viruses that affect livestock worldwide. The 1B capsid of FMDV has been widely used to detect and confirm the infection. In the present study, the sequence coding for 1B subunit of FMDV capsid was expressed in insect cells using the baculovirus expression system under the polyhedrin (polh) promoter. The expression of 1B capsid protein was validated in the culture filtrate of insect cells using SDS-PAGE and western blotting. The culture filtrate containing recombinant 1B capsid (r1B) was used as a coated antigen in an indirect enzyme-linked immunosorbent assay (ELISA). The antigenicity and specificity of r1B against SAT 2 serotype-specific antibodies were assessed. Our results revealed that a protein concentration as low as 25 ng could detect SAT 2-specific antibodies in ELISA. The results highlight the application of insect cells developed r1B protein in the detection of FMDV. Further studies are required to determine the ability of r1B to detect other FMDV serotypes.

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“…This difference was attributed to posttranslational modifications, principally N-glycosylation, the rgE amino acid sequence contained several potential glycosylation sites and disulphide bonds (data not shown). Furthermore, there are reports of rgE with an apparent Mm of~80 kDa when expressed in a Baculovirus-insect system [34,36].…”

Section: Cloning and Expression Of Rge In P Pastorismentioning

confidence: 99%

Secretory expression of bovine herpesvirus type 1/5 glycoprotein E in Pichia pastoris for the differential diagnosis of vaccinated or infected cattle

Siedler

Roloff

Sá

et al. 2017

Protein Expression and Purification

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Bovine herpesvirus (BoHV) glycoprotein E (gE) is a non-essential envelope glycoprotein and the deletion of gE has been used to develop BoHV-1 and BoHV-5 differential vaccine strains. The DIVA (Differentiation of Infected from Vaccinated Animals) strategy, using marker vaccines based on gE-negative BoHV strains, allows the identification of vaccinated or infected animals in immunoassays designed to detect anti-gE antibodies. In this study a codon optimized synthetic sequence of gE containing highly conserved regions from BoHV-1 and BoHV-5 was expressed in Pichia pastoris. Following expression, the recombinant gE (rgE) was secreted and purified from the culture medium. The rgE was identified by Western blotting (WB) using sera from cattle naturally infected with BoHV-1 and/or BoHV-5, or sera from bovines experimentally infected with wild-type BoHV-5. Sera collected from cattle vaccinated with a BoHV-5 gI/gE/US9¯ marker vaccine failed to recognise rgE. Expression of rgE, based on a sequence containing highly conserved regions from BoHV-1 and BoHV-5, in P. pastoris enabled the production of large quantities of rgE suitable for use in immunoassays for the differentiation vaccinated or infected cattle.

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Baculovirus expression and diagnostic utility of the glycoprotein E of bovine herpesvirus-1.1 Egyptian strain “Abu-Hammad”

Cited by 7 publications

References 19 publications

Use of baculovirus expression system for generation of virus-like particles: Successes and challenges

Use of baculovirus expression system for generation of virus-like particles: Successes and challenges

Expression of 1B capsid protein of Foot-and-mouth disease virus (FMDV) using baculovirus expression system and its validation in detecting SAT 2- specific antisera

Secretory expression of bovine herpesvirus type 1/5 glycoprotein E in Pichia pastoris for the differential diagnosis of vaccinated or infected cattle

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