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Biologics



biologics@ibet.pt

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iBET’s Bioproduction Unit offers R&D services for virus-based biopharmaceuticals production

With 30 years of experience and equipped with state-of-the-art equipment, iBET is the ideal partner for production and purification of replicative virus and non-replicative viral particles

FULL RANGE SERVICE: FROM GENE TO PROTEIN

iBET’s Bioproduction Unit offers late-stage R&D, scale up and technology transfer services, from small-scale feasibility studies to large-scale production and purification, and technology transfer to GMP (provided through GenIbet, www.genibet.com). At iBET a full set of instruments and strategies have been established to develop integrated bioprocesses and analytical methods for the production, purification and characterization of virus-based biopharmaceuticals. The infrastructure is located in a BSL2 facility allowing scale-up to 50 L.

Virus-based pharmaceuticals

Upstream processing

  • Cell lines:
    • Mammalian cells: HEK293, PerC6, CAP, Caco2, MRC-5, Sp2/0, BAE, Vero, MDCK, DK, CHO, A549, ExpiCHO-S™ and Expi293F™;
    • Insect cells: Sf-9, Sf-21, High Five, ExpiSf9™ and super Sf9.
  • Expression systems:
    • Stable expression in mammalian and insect cells;
    • Transient expression, e.g. transfection and insect cells-baculovirus expression vector system (IC-BEVS).
  • Culture systems:
    • Anchorage dependent cells, e.g. microcarrier technology in bioreactors;
    • Single cell suspension.
  • Bioreactor systems/types and operation modes:
    • Stainless-steel and single-use bioreactors;
    • Environmentally controlled stirred tank (0.5 L to 50 L) and Wave bioreactors (0.5 L to 25 L);
    • Batch, Fed-batch, perfusion and continuous.

Downstream processing

  • Purification of secreted and intracellular viral particles;
  • Ultracentrifugation;
  • Membrane technology: ÄKTA Crossflow – scalable, ultrafiltration systems;
  • Chromatography: ÄKTA Platform Technology (Pure, Avant and Pilot);
  • Continuous and multi-column chromatographic purification systems.

Product Characterization

  • Impurities: DNA, Host Cell Protein, product related impurities (e.g. empty capsids);
  • Titration protocols for infectious viral particles, e.g. flow cytometry, TCID50, plaque assay and MTT assay;
  • RT-qPCR for genome-containing viral particles and for adventitious viruses;
  • Capillary Electrophoresis;
  • ELISA for transgene expression and for process contaminant detection;
  • Nanoparticle tracking analysis (using NanoSight®) for physical viral particles quantification;
  • Mass spectrometry;
  • Transmission electron microscopy (negative staining and immunogold labelling);
  • Dynamic light scattering;
  • Surface plasmon resonance;
  • Bio-layer interferometry technology;
  • Tunable Resistive Pulse Sensing (qNANO);
  • Analytical chromatographic methods for purity assessment;
  • Cell-based potency assays.

Associated Services

At iBET, we develop mammalian and insect cell lines for stable expression of virus-based biopharmaceuticals using advanced molecular biotechnology and synthetic biology tools (e.g. recombinase mediated cassette exchange (RMCE), metabolic engineering and CRISPR-CAS9). We also develop computational biology methods (e.g. dielectric and fluorescence spectroscopy, holographic imaging) for on-line monitoring of cell concentration and viral particles production.

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