Quality control and analytical methods for baculovirus-based products

Roldão, António; Vicente, Tiago; Peixoto, Cristina; Carrondo, Manuel J.T.; Alves, Paula M.

doi:10.1016/j.jip.2011.05.009

Cited by 19 publications

(26 citation statements)

References 111 publications

(108 reference statements)

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“…However, the expression efficiency of foreign proteins using the polyhedrin promoter could not obtain the protein yields observed for native polyhedrin. As a result of ongoing studies and efforts over the last decade, BEVS has evolved to overcome some of these technical issues [11], [12]. Many researchers have performed studies to resolve this limitation, including the alteration of promoter sequences, fusion expression with partial polyhedrin or various tagging signals and co-expression with regulatory proteins[13]–[18].…”

Section: Introductionmentioning

confidence: 99%

Hyper-Enhanced Production of Foreign Recombinant Protein by Fusion with the Partial Polyhedrin of Nucleopolyhedrovirus

et al. 2013

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To enhance the production efficiency of foreign protein in baculovirus expression systems, the effects of polyhedrin fragments were investigated by fusion expressing them with the enhanced green fluorescent protein (EGFP). Recombinant viruses were generated to express EGFP fused with polyhedrin fragments based on the previously reported minimal region for self-assembly and the KRKK nuclear localization signal (NLS). Fusion expressions with polyhedrin amino acids 19 to 110 and 32 to 110 lead to localization of recombinant protein into the nucleus and mediate its assembly. The marked increase of EGFP by these fusion expressions was confirmed through protein and fluorescence intensity analyses. The importance of nuclear localization for enhanced production was shown by the mutation of the NLS within the fused polyhedrin fragment. In addition, when the polyhedrin fragment fused with EGFP was not localized in the nucleus, some fragments increased the production of protein. Among these fragments, some degradation of only the fused polyhedrin was observed in the fusion of amino acids 19 to 85 and 32 to 85. The fusion of amino acids 32 to 85 may be more useful for the enhanced and intact production of recombinant protein. The production of E2 protein, which is a major antigen of classical swine fever virus, was dramatically increased by fusion expression with polyhedrin amino acids 19 to 110, and its preliminary immunogenicity was verified using experimental guinea pigs. This study suggests a new option for higher expression of useful foreign recombinant protein by using the partial polyhedrin in baculovirus.

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Section: Introductionmentioning

confidence: 99%

Hyper-Enhanced Production of Foreign Recombinant Protein by Fusion with the Partial Polyhedrin of Nucleopolyhedrovirus

et al. 2013

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“…VLPs are classified in two types based on structure: enveloped VLPs, that include a host-derived lipid membrane and naked or nonenveloped (simply referred to as VLPs). Likewise, they are also classified as simple or multiple VLPs depending on the number of different recombinant proteins included in the structure (Noad and Roy, 2003;Roldão et al, 2011). Some difficulties may be associated with VLP production: for example, protein stoichiometry can be a problem since the incorporation of adequate proportions of each protein may not occur (Latham and Galarza, 2001).…”

Section: The Baculovirus Expression Vector System For the Generation mentioning

confidence: 99%

Baculovirus-Derived Vectors for Immunization and Therapeutic Applications

Fabre

Arrías

Masson

et al. 2020

Emerging and Reemerging Viral Pathogens

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“…Thus, an effective downstream process for viral vectors needs to be established, which efficiently removes impurities while keeping the viral particles intact (see also Roldão et al in 2011). The commonly used purification and concentration methods for baculovirus vectors are based on ultracentrifugation (Airenne et al, 2000), but this method is labor-intensive, time consuming and the scalability is limited.…”

Section: Upstream and Downstream Process For Baculoviral Vectorsmentioning

confidence: 99%

“…The consistency of the production process should be determined by producing at least three batches of the bulk product characterized for identity, purity, potency and safety (The European Agency for the Evaluation of Medicinal Products, 2001). Relevant articles by Dalemans (2006), and Cox and Hollister (2009) describe the characterization of baculovirus related material for vaccine production (for a more comprehensive review about quality control of baculoviruses, see Roldão et al, 2011).…”

Section: Characterization Of the Bv Productmentioning

confidence: 99%