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António Roldão

Head of Lab

Health & Pharma Division
TCA - Science & Services

+ 351 21 446 9 418

Explore Innovation

Explore Innovation

Bioprocess R&D and analytics for vaccines production has been an area of research at iBET for more than 20 years

We have expertise in subunit vaccines, including virus-like particles (VLPs) and recombinant proteins, vectored vaccines and live attenuated and inactivated vaccines. The exploitation of VLPs as platforms for chimeric antigen presentation is also being conducted at iBET

iBET’s Cell-based Vaccines Development lab is focused on early-stage bioprocess R&D (from molecular biology to small-scale expression and purification), process monitoring and control, product characterization and purity assessment. All activities are carried out in BSL2 labs and certified according to GLP standards.

Portfolio of candidate vaccines

Human Vaccines

  • Non-enveloped and enveloped virus-like particles (VLPs) (e.g. Influenza, Dengue, Rotavirus, HIV);
  • Chimeric VLPs (e.g. HCV retroVLPs);
  • Recombinant (chimeric) proteins (e.g. Malaria, Chagas, SARS-CoV-2).

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).

Molecular Biology

iBET develops mammalian and insect cell lines for stable expression of vaccine candidates using advanced molecular biotechnology and synthetic biology tools.

  • Classical molecular biology tools for cell and virus genetic manipulation;
  • Recombinase mediated cassette exchange (RMCE) technology for stable cell line development;
  • Knock down and knockout systems (e.g. CRISP/Cas9).

Upstream Processing

iBET is focused on devising strategies to monitor, control and optimize product titer and quality of vaccines.

Cell lines, viruses and expression systems

  • 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);
    • Transient (insect cells-baculovirus expression system or transfection) and stable expression (RMCE technology).

Culture systems

  • Anchorage dependent cells – (i) microcarrier technology in bioreactors and (ii) roller bottles and cell factories;
  • Single cell suspension and cell aggregate based (microcarrier-free) approaches in bioreactors.

Bioreactor systems/types and operation modes

  • Stainless-steel and single-use bioreactors;
  • Environmentally controlled stirred tank (0.25-50 L) and Wave bioreactors (0.5-25 L);
  • Batch, Fed-batch, perfusion (using XCell™ ATF System technology) and continuous.

Downstream processing

iBET targets innovative solutions for increasing product recovery yields and quality while reducing manufacturing costs.

  • Chromatographic purification using ÄKTA platform (Prime, Explorer, Pure,  Avant and Pilot);
  • End-to-end single-use purification platforms;
  • Ultra and microfiltration devices (from lab to preparative scale);
  • Expanded bed adsorption and radial-flow chromatography;
  • Continuous and multi-column chromatographic purification systems;
  • Scaled-down devices for optimization of chromatographic adsorption and elution conditions;
  • Ion exchange, size exclusion, affinity and hydrophobic interaction chromatographic purification with membrane and resin technology (from lab to preparative scale).

Process monitoring and control

iBET develops mathematical models and integrated systems for detailed bioprocess characterization, on-line bioprocess monitoring and control. In addition, omics technologies are applied to guide rational feeding strategies towards improved productivities.

  • Integrated systems for on-line bioprocess monitoring and control (e.g. 2D-fluorometry);
  • UBICON system for process control and supervision (the basis for cGMP dossiers);
  • Deterministic, stochastic and hybrid mathematical models based on mass transfer balances and thermodynamics;
  • Bioconjugate chemistry for virus labelling and bioprocess monitoring;
  • Transcriptomics (microarray analysis and RNAseq);
  • Proteomics (MS, Thermofluor, Limited Proteolysis technology, X-ray crystallography, NMR);
  • Metabolomics (HPLC, 13C/1H-NMR and GC-MS);
  • Fluxomics (classical MFA, 13C-MFA and metabolic pathway analysis).

Product characterization and purity assessment

iBET has extended expertise in developing in process assays for characterisation of product, cells and host-virus interactions, in particular assays developed and qualified for technology transfer.

  • Titration protocols for infectious viral particles (e.g. flow cytometry, TCID50, plaque assay);
  • Real time qPCR for genome-containing viral particles and for adventitious viruses;
  • Capillary electrophoresis;
  • ELISA for transgene expression and for process contaminant detection;
  • Mass spectrometry;
  • Transmission electron microscopy (negative staining and immunogold labelling);
  • Surface plasmon resonance (Biacore);
  • Biolayer interferometry analysis (Octet);
  • Nanoparticle tracking analysis (using NanoSight®);
  • Tunable resistive pulse sensing (qNANO);
  • Dynamic light scattering;
  • Analytical size-exclusion for purity assessment;
  • Cell-based potency assays.

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