A Novel Bacterial Protease Inhibitor Adjuvant in RBD-Based COVID-19 Vaccine Formulations Containing Alum Increases Neutralizing Antibodies, Specific Germinal Center B Cells and Confers Protection Against SARS-CoV-2 Infection in Mice

Coria, Lorena M.; Saposnik, Lucas M; Castro, Celeste Pueblas; Castro, Eliana; Bruno, Luciana; Stone, William B.; Pérez, Paula Soledad; Darriba, M. Laura; Chemes, Lucía B.; Alcaín, Julieta; Mazzitelli, Ignacio; Varese, Augusto; Salvatori, Melina; Auguste, Albert J.; Álvarez, Diego E.; Pasquevich, Karina A.; Cassataro, Juliana

doi:10.3389/fimmu.2022.844837

Cited by 15 publications

(15 citation statements)

References 69 publications

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“…Also, the bioprocess can be easily scalable without affecting the productivity and the protein quality (O’Flaherty et al 2020 ). The majority of the reports published to date describing the expression of RBD domain in HEK293 cells choose the transient production instead of the stable cell line generation (Stadlbauer et al 2020 ; Castro et al 2021 ; Mehdi et al 2021 ; Mehalko et al 2021 ; Coria et al 2022 ; Gstöttner et al 2022 ; Amanat et al 2020 ).…”

Section: Discussionmentioning

confidence: 99%

Design and optimization of an IgG human ELISA assay reactive to recombinant RBD SARS-CoV-2 protein

Rodríguez

Ceaglio

Gugliotta

et al. 2022

Appl Microbiol Biotechnol

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Serology assays are essential tools to mitigate the effect of COVID-19, help to identify previous SARS-CoV-2 infections or vaccination, and provide data for surveillance and epidemiologic studies. In this study, we report the production and purification process of the receptor-binding domain (RBD) of SARS-CoV-2 in HEK293 cells, which allowed the design, optimization, and validation of an indirect ELISA (iELISA) for the detection of human anti-RBD antibodies. To find the optimal conditions of this iELISA, a multivariate strategy was performed throughout design of experiments (DoE) and response surface methodology (RSM), one of the main tools of quality by design (QbD) approach. The adoption of this strategy helped to reduce the time and cost during the method development stage and to define an optimum condition within the analyzed design region. The assay was then validated, exhibiting a sensitivity of 94.24 (86.01–98.42%; 95% CI) and a specificity of 95.96% (89.98–98.89%; 95% CI). Besides, the degree of agreement between quality results assessed using kappa’s value was 0.92. Hence, this iELISA represents a high-throughput technique, simple to perform, reliable, and feasible to be scaled up to satisfy the current demands. Since RBD is proposed as the coating antigen, the intended use of this iELISA is not only the detection of previous exposure to the virus, but also the possibility of detecting protective immunity. Key points • RBD was produced in 1-L bioreactor and highly purified. • An iELISA assay was optimized applying QbD concepts. • The validation procedure demonstrated that this iELISA is accurate and precise. Graphical Abstract Supplementary Information The online version contains supplementary material available at 10.1007/s00253-022-12254-w.

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Section: Discussionmentioning

confidence: 99%

Design and optimization of an IgG human ELISA assay reactive to recombinant RBD SARS-CoV-2 protein

Rodríguez

Ceaglio

Gugliotta

et al. 2022

Appl Microbiol Biotechnol

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“…Once both DNA and RNA vaccines have limitations regarding their therapeutic effects, such as a modest percentage of regression or cure, a low correlation between tumor regression and IFNγ responses, and limited thermostability [61,76,77], some strategies to improve T cell function may be necessary to achieve optimal immune responses [36,39]. Among these optimizations are, for example, constructs containing synthetic antigens with multiple epitopes for TC8+, TCD4+, or both, the optimization of codons in the sequences of DNA and RNA, in addition to the use of fat-soluble delivery systems, and platforms with natural adjuvants in the cell wall [80]. The next topic addresses delivery systems used for nucleic acid vaccines that could be further employed in future HPV vaccine strategies.…”

Section: Nucleic Acid Vaccines Against Hpv-related Cancers and Their ...mentioning

confidence: 99%

“…However, although these vectors can induce robust immune responses, some of them, especially viruses and bacteria, can be neutralized by components of the host hu-moral immunity after repeated administration and exhibit some cytotoxicity [80].…”

Section: Microorganism-based Delivery Systemsmentioning

confidence: 99%

“…First, the use of live bacteria, albeit in modified or attenuated form, includes the likelihood of causing infection, particularly in infants and immunocompromised patients [176]. Second, bacterial delivery systems, such as viruses, develop neutralizing antibodies against the bacteria (vector) itself, resulting in the decreased efficacy of the vaccine antigen [80].…”

Section: Archaea Bacteria and Their By-productsmentioning

confidence: 99%

“…Major safety concerns associated with the various delivery systems are biocompatibility, biodistribution, and the induction of unwanted immune reactions [51]. For example, the repeated application of adenoviral, bacterial, and PLL-and PAMAM-based systems can neutralize the carrier molecule and not the target antigen [80,114]. In therapeutic vaccines that require larger applications at shorter intervals, this unrequited neutralization may interfere with the immune response.…”

Section: The Influence Of Delivery Systems On the Immune Responsementioning

confidence: 99%

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Enhancing the Effect of Nucleic Acid Vaccines in the Treatment of HPV-Related Cancers: An Overview of Delivery Systems

et al. 2022

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Prophylactic vaccines against human papillomavirus (HPV) have proven efficacy in those who have not been infected by the virus. However, they do not benefit patients with established tumors. Therefore, the development of therapeutic options for HPV-related malignancies is critical. Third-generation vaccines based on nucleic acids are fast and simple approaches to eliciting adaptive immune responses. However, techniques to boost immunogenicity, reduce degradation, and facilitate their capture by immune cells are frequently required. One option to overcome this constraint is to employ delivery systems that allow selective antigen absorption and help modulate the immune response. This review aimed to discuss the influence of these different systems on the response generated by nucleic acid vaccines. The results indicate that delivery systems based on lipids, polymers, and microorganisms such as yeasts can be used to ensure the stability and transport of nucleic acid vaccines to their respective protein synthesis compartments. Thus, in view of the limitations of nucleic acid-based vaccines, it is important to consider the type of delivery system to be used—due to its impact on the immune response and desired final effect.

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A bioengineered pseudovirus nanoparticle displaying SARS‐CoV 2 RBD fully protects mice from mortality and weight loss caused by SARS‐CoV 2 challenge

Xia

López

Vago³

et al. 2023

Biotechnology Journal

Self Cite

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The COVID‐19 pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) has caused considerable morbidity and mortality worldwide. Although authorized COVID‐19 vaccines have been shown highly effective, their significantly lower efficacy against heterologous variants, and the rapid decrease of vaccine‐elicited immunity raises serious concerns, calling for improved vaccine tactics. To this end, a pseudovirus nanoparticle (PVNP) displaying the receptor binding domains (RBDs) of SARS‐CoV‐2 spike, named S‐RBD, was generated and shown it as a promising COVID‐19 vaccine candidate. The S‐RBD PVNP was produced using both prokaryotic and eukaryotic systems. A 3D structural model of the S‐RBD PVNPs was built based on the known structures of the S60 particle and RBDs, revealing an S60 particle‐based icosahedral symmetry with multiple surface‐displayed RBDs that retain authentic conformations and receptor‐binding functions. The PVNP is highly immunogenic, eliciting high titers of RBD‐specific IgG and neutralizing antibodies in mice. The S‐RBD PVNP demonstrated exceptional protective efficacy, and fully (100%) protected K18‐hACE2 mice from mortality and weight loss after a lethal SARS‐CoV‐2 challenge, supporting the S‐RBD PVNPs as a potent COVID‐19 vaccine candidate. By contrast, a PVNP displaying the N‐terminal domain (NTD) of SARS‐CoV‐2 spike exhibited only 50% protective efficacy. Since the RBD antigens of our PVNP vaccine are adjustable as needed to address the emergence of future variants, and various S‐RBD PVNPs can be combined as a cocktail vaccine for broad efficacy, these non‐replicating PVNPs offer a flexible platform for a safe, effective COVID‐19 vaccine with minimal manufacturing cost and time.

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A Novel Bacterial Protease Inhibitor Adjuvant in RBD-Based COVID-19 Vaccine Formulations Containing Alum Increases Neutralizing Antibodies, Specific Germinal Center B Cells and Confers Protection Against SARS-CoV-2 Infection in Mice

Cited by 15 publications

References 69 publications

Design and optimization of an IgG human ELISA assay reactive to recombinant RBD SARS-CoV-2 protein

Design and optimization of an IgG human ELISA assay reactive to recombinant RBD SARS-CoV-2 protein

Enhancing the Effect of Nucleic Acid Vaccines in the Treatment of HPV-Related Cancers: An Overview of Delivery Systems

A bioengineered pseudovirus nanoparticle displaying SARS‐CoV 2 RBD fully protects mice from mortality and weight loss caused by SARS‐CoV 2 challenge

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