Vaccines Bioprocess Engineering

Integrating multidisciplinary cutting-edge technologies to create innovative bioprocesses for production of vaccines.

Vaccines Bioprocess Engineering at iBET

In the rapidly evolving field of vaccine development, iBET leverages cutting-edge research and innovative technologies to address the most pressing challenges: ensuring efficacy, safety, bioprocess robustness and scalability, for both veterinary and human vaccines.

Our team has over 25 years of expertise in early-stage bioprocess R&D (from molecular biology to small-scale expression and purification) for vaccine production based on mammalian and insect cells. Our bespoke approach additionally includes process monitoring and control and bioanalytics for product characterization and purity assessment.

iBET’s vaccine portfolio expertise includes subunit vaccines (virus-like particles (VLPs) and recombinant proteins), vectored vaccines, and live attenuated vaccines. We also exploit VLPs as platforms for chimeric antigen presentation. All activities are carried out in BSL2 labs.

Portfolio of candidate vaccines

Human Vaccines

• Non-enveloped and enveloped virus-like particles (VLPs) (e.g. Influenza, Dengue, Rotavirus, HIV).
• Chimeric VLPs (e.g. retroVLPs functionalized with HCV E1/E2, ferritin nanoparticles functionalized with Rift Valley Fever Gn or SARS-CoV-2 RBD).
• Recombinant (chimeric) proteins (e.g. Malaria, Chagas, SARS-CoV-2 S and RBD).

Veterinary Vaccines

• Non-enveloped VLPs (e.g. Porcine Parvovirus, RHDV).
• Live attenuated (e.g. Peste des Petits Ruminants Virus – PPRV).
• Recombinant adenovirus-based (e.g. Rinderpest and PPRV).

Related Technologies

Cell Line Development

Establishment of mammalian and insect cell lines for stable expression of vaccine candidates using advanced molecular biotechnology and synthetic biology tools, e.g. recombinase mediated cassette exchange (RMCE) technology for stable cell line development and knock-in and -out systems (e.g. CRISPR/Cas9).

Expression Systems

Our vaccine production platforms are based on mammalian cells (e.g. Vero, MRC 5, BHK, CHO, HEK293), insect cells (e.g. Sf-9, Sf-21 and High Five), bacteria (e.g. E. coli) and yeast (e.g. P. Pastoris). Production of our vaccine candidates can be done using transient (e.g. insect cells-baculovirus expression system (IC-BEVS), transfection in mammalian cells) or stable (i.e. using RMCE technology in both mammalian and insect cells) expression systems.

Upstream Processing

We employ our extensive know-how and expertise in cell culture systems for bioprocess development and intensification. Our state-of-the-art culture systems comprise glass and single-use stirred-tank bioreactors (0.5 to 5L). Bioreactor operation modes used include batch, fed-batch, perfusion and continuous.

Downstream Processing

We target innovative solutions for increasing product recovery yields and quality. Current purification approaches combine scalable membrane and chromatography-based technologies (e.g. using ÄKTA platform) with filtration-based strategies (ultra and microfiltration systems, dead-end filtration and tangential flow filtration using hollow fibers and flat sheet cassettes). Integrated continuous purification approaches are also available using continuous or multi-column chromatographic purification systems tailored to the product features.

Bioprocess monitoring and control

We develop mathematical models (deterministic, stochastic and hybrid) and integrated systems for detailed bioprocess characterization, on-line bioprocess monitoring and control (e.g. 2D fluorometry). In addition, omics technologies, such as transcriptomics, proteomics metabolomics and fluxomics are applied to guide rational feeding strategies towards improved productivities.

Bioanalytics for product characterization

Development of in-process assays for product characterisation, suitable for technology transfer. Technologies used include cell-based potency assays, capillary electrophoresis-based methods, nanoparticle tracking analysis, dynamic light scattering, mass photometry and surface plasmon resonance based-methods. The product’s thermal and aggregation stability is characterized using different biophysical methodologies such as differential scanning fluorimetry and thermal shift assay.

Highlights

Snakebite antivenom based on virus-like particles

iBET participates in EU-funded “ADDovenom” project (EU-FET) aiming to create a unique type of snakebite antivenom treatment to neutralize and eliminate venom toxins with enhanced efficacy, safety and affordability when compared to treatments currently available.

iBET’s role is to establish a scalable, GMP-compliant platform for production of customized ADDomers as snakebite antivenoms. A white paper reviewing the project’s goals, experimental strategies and expected outcomes of the project was recently published (Menzies et al 2023).

ADDovenom Project (EU-FET)

New malaria (PfRipr5/SA-1) vaccine candidate

The Japanese GHIT Fund is supporting the development of a new asexual-blood stage malaria vaccine by Ehime University and Sumitomo Pharma Co., Ltd. (Japan), European Vaccine Initiative e.V. (Germany), and iBET.

iBET’s role in the PfRipr5-PD project is to develop a scalable and cGMP-compliant process for manufacturing PfRipr5 for safety-toxicology studies. Some of the scientific results from this project have been published by Correia et al (2022) and Takashima et al (2022).

PfRipr5-PD Project (JP-GHIT)

Bioprocess intensification for continuous production of influenza VLPs

A multi-stage bioreactor design was established for continuous production of influenza VLPs using the insect cell-baculovirus expression vector system (IC-BEVS), expanding the applicability of IC-BEVS for process intensification approaches.

Tyrosinase-mediated bioconjugation

Within EU-funded “ProMeTeus” project (MSCA-RISE-2018), ferritin nanoparticles were functionalized with Rift Valley Fever Gn or SARS-CoV-2 RBD through tyrosinase-mediated bioconjugation. This innovative modular approach for vaccine development holds promise for timely and effective responses to emerging infectious threats.

ProMeTeus Project (MSCA-RISE-2018)

Building a virosome-based COVID-19 vaccine candidate

Within “TRANSVAC2” project (H2020-INFRAIA) and in collaboration with Mymetics BV (The Netherlands), we established a scalable bioprocess to produce high-quality SARS-CoV-2 Spike protein using the insect cell-baculovirus expression vector system for inclusion in a virosome-based COVID-19 vaccine candidate.

Published work related to this project is available here.

TRANSVAC2 Project (H2020-INFRAIA)

Single cell RNA sequencing in insect cells

Within “STACCATO” project (H2020-MSCA-ITN) and in collaboration with NIBRT (Ireland), we implemented a scRNA-seq pipeline to analyse transcriptomic signatures in insect (Sf9 and High Five) cells during the production of influenza virus-like particles (as vaccine candidate) and adeno-associated virus (for gene therapy), using the baculovirus expression vector system.

Published work related to this project can be found in the following publications: N. Virgolini 2023, N. Virgolini II 2023, M. Silvano 2023, M. Silvano 2022.

STACCATO Project (H2020-MSCA-ITN)

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Related Teams

Cell-Based Vaccines Development Lab

António Roldão

Head of the Cell-based Vaccines Development Laboratory & Coordinator of Late-Stage R&D Unit

Multidisciplinary cutting-edge technologies to develop innovative bioprocesses for vaccine production

Downstream Process Development Lab

Cristina Peixoto

Head of Downstream Process Development lab, Coordinator of Bioproduction Unit

Ground-breaking technologies for purification of new modalities

Cell Line Development and Molecular Virology Lab

Ana Sofia Coroadinha

Head of Cell Line Development and Molecular Virology Lab

Development and engineering of virus-based biopharmaceuticals for gene therapy and vaccines.

Late-Stage R&D and Bioproduction Unit Team