Chrysalises as natural production units for recombinant subunit vaccines

Escribano, José M.; Cid, Miguel; Reytor, Edel; Alvarado, Carmen; Núñez, María; Martínez-Pulgarín, Susana; Dalton, Romy M.

doi:10.1016/j.btecx.2020.100019

Cited by 9 publications

(18 citation statements)

References 24 publications

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“…The production of SARS-CoV-2 N protein in insect pupae ( Tricoplusia ni ; T. ni ) was performed as previously described ( Escribano et al, 2020 ). Briefly, pupae were allocated in the inoculation robot that dispensed a maximum of 5 μl with the baculovirus titers protein in 5 days pupae incubation time in constant temperature and humidity chambers.…”

Section: Methodsmentioning

confidence: 99%

Identification of Niemann-Pick C1 protein as a potential novel SARS-CoV-2 intracellular target

et al. 2021

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Niemann-Pick type C1 (NPC1) receptor is an endosomal membrane protein that regulates intracellular cholesterol traffic. This protein has been shown to play an important role for several viruses. It has been reported that SARS-CoV-2 enters the cell through plasma membrane fusion and/or endosomal entry upon availability of proteases. However, the whole process is not fully understood yet and additional viral/host factors might be required for viral fusion and subsequent viral replication. Here, we report a novel interaction between the SARS-CoV-2 nucleoprotein (N) and the cholesterol transporter NPC1. Furthermore, we have found that some compounds reported to interact with NPC1, carbazole SC816 and sulfides SC198 and SC073, were able to reduce SARS-CoV-2 viral infection with a good selectivity index in human cell infection models. These findings suggest the importance of NPC1 for SARS-CoV-2 viral infection and a new possible potential therapeutic target to fight against COVID-19.

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Section: Methodsmentioning

confidence: 99%

Identification of Niemann-Pick C1 protein as a potential novel SARS-CoV-2 intracellular target

et al. 2021

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“…Yield of expression of up to 2 mg of recombinant protein per larvae has been reported for T. ni as expression system [262]. Protein expression in T. ni pupae has also emerged as an innovative platform and expression of several proteins have been reported, reaching productivities on the range of milligrams per infected pupa [265]. As with transgenic animals, a societal debate also comes up regarding the use of biotechnology in insects; however, this platform allows for heterologous protein expression with no need to genetically modify the host genome.…”

Section: Insectsmentioning

confidence: 99%

Platforms for Production of Protein-Based Vaccines: From Classical to Next-Generation Strategies

Cid¹,

Bolı́var

2021

Biomolecules

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To date, vaccination has become one of the most effective strategies to control and reduce infectious diseases, preventing millions of deaths worldwide. The earliest vaccines were developed as live-attenuated or inactivated pathogens, and, although they still represent the most extended human vaccine types, they also face some issues, such as the potential to revert to a pathogenic form of live-attenuated formulations or the weaker immune response associated with inactivated vaccines. Advances in genetic engineering have enabled improvements in vaccine design and strategies, such as recombinant subunit vaccines, have emerged, expanding the number of diseases that can be prevented. Moreover, antigen display systems such as VLPs or those designed by nanotechnology have improved the efficacy of subunit vaccines. Platforms for the production of recombinant vaccines have also evolved from the first hosts, Escherichia coli and Saccharomyces cerevisiae, to insect or mammalian cells. Traditional bacterial and yeast systems have been improved by engineering and new systems based on plants or insect larvae have emerged as alternative, low-cost platforms. Vaccine development is still time-consuming and costly, and alternative systems that can offer cost-effective and faster processes are demanding to address infectious diseases that still do not have a treatment and to face possible future pandemics.

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“…Trichoplusia ni (T. ni; Cabbage looper) insects were reared following a previously described methodology [35]. Briefly, eggs were placed into larvae developmental cages, containing an artificial insect diet.…”

Section: Production Of Recombinant Proteins In Insect Pupaementioning

confidence: 99%

“…However, to date, none of the vaccines targeting RHDV have been successfully commercialised. We therefore aimed to develop a dual RHDVGI.1/RHDVGI.2 vaccine using the Baculovirus Expression Vector System (BEVS) using CrisBio technology, based on the use of Trichoplusia ni pupae as natural bioreactors [35] to increase vaccine production efficiency in a cost-effective manner. In order to reduce the downstream production-associated costs, our strategy was to generate chimeric VLPs containing the VP60 proteins from the two serotypes obtained by co-infection of the insect pupae with two recombinant baculoviruses expressing either capsid protein.…”

Section: Introductionmentioning

confidence: 99%

Chimeric VLPs Bearing VP60 from Two Serotypes of Rabbit Haemorrhagic Disease Virus Are Protective against Both Viruses

Dalton

Alvarado²,

Reytor³

et al. 2021

Vaccines

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The VP60 capsid protein from rabbit haemorrhagic disease virus (RHDV), the causative agent of one of the most economically important disease in rabbits worldwide, forms virus-like particles (VLPs) when expressed using heterologous protein expression systems such as recombinant baculovirus, yeasts, plants or mammalian cell cultures. To prevent RHDV dissemination, it would be beneficial to develop a bivalent vaccine including both RHDV GI.1- and RHDV GI.2-derived VLPs to achieve robust immunisation against both serotypes. In the present work, we developed a strategy of production of a dual-serving RHDV vaccine co-expressing the VP60 proteins from the two RHDV predominant serotypes using CrisBio technology, which uses Tricholusia ni insect pupae as natural bioreactors, which are programmed by recombinant baculovirus vectors. Co-infecting the insect pupae with two baculovirus vectors expressing the RHDV GI.1- and RHDV GI.2-derived VP60 proteins, we obtained chimeric VLPs incorporating both proteins as determined by using serotype-specific monoclonal antibodies. The resulting VLPs showed the typical size and shape of this calicivirus as determined by electron microscopy. Rabbits immunised with the chimeric VLPs were fully protected against a lethal challenge infection with the two RHDV serotypes. This study demonstrates that it is possible to generate a dual cost-effective vaccine against this virus using a single production and purification process, greatly simplifying vaccine manufacturing.

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Chrysalises as natural production units for recombinant subunit vaccines

Cited by 9 publications

References 24 publications

Identification of Niemann-Pick C1 protein as a potential novel SARS-CoV-2 intracellular target

Identification of Niemann-Pick C1 protein as a potential novel SARS-CoV-2 intracellular target

Platforms for Production of Protein-Based Vaccines: From Classical to Next-Generation Strategies

Chimeric VLPs Bearing VP60 from Two Serotypes of Rabbit Haemorrhagic Disease Virus Are Protective against Both Viruses

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