iBET PhD Portraits – April/May 2022

Welcome to the first article of the new series "iBET PhD portraits", where we will showcase and celebrate the conclusion of PhD Thesis Projects at iBET.

iBET has three newly-graduated PhDs this Spring: Bárbara Fernandes (Cell-Based Vaccines Development Lab), Ricardo Correia (Cell-Based Vaccines Development Lab) and Jorge Moura-Sampaio (Novartis Pharma Lab).

Congratulations to Bárbara, Ricardo, and Jorge for this important achievement in your scientific careers and best wishes for your new positions at iBET.

Find below a PhD portrait of each of our new graduates.

Ricardo Correia

PhD in Bioengineering, ITQB NOVA, NOVA University Lisbon (27^th April 2022)

Thesis Title: Atypical culture conditions and process intensification to enhance vaccine production using insect cells

Thesis Highlights:

• Improved production of influenza VLPs using adaptive laboratory evolution of insect High Five cells to neutral pH;
• Enhanced storage stability of influenza VLPs using a trehalose-glycerol natural deep eutectic solvent solution;
• Improved production of the malaria vaccine candidate PfRipr5 through low culture temperature and baculovirus vector optimization;
• Establishment of a continuous multi-stage bioreactor process to produce influenza VLPs;

Publications:

• R Correia et al (2021) Improved storage of influenza HA-VLPs using a trehalose-glycerol natural deep eutectic solvent system. Vaccine 39(24):3279-3286.

• R Correia et al (2020) Improving Influenza HA-Vlps Production in Insect High Five Cells via Adaptive Laboratory Evolution. MDPI Vaccines 8(4), 589

• D Sequeira et al (2018) Combining stable insect cell lines with baculovirus-mediated expression for multi-HA influenza VLP production. Vaccine 36(22): 3112-3123.

New Position: Research Associate at the Cell-Based Vaccines Development Lab (50%) and in a Service Project for Turnstone Biologics, Inc (50%), iBET.

Bárbara Fernandes

PhD in Molecular Biosciences, ITQB NOVA, NOVA University Lisbon (29^th April 2022)

Thesis Title: Bioprocess Engineering of Insect Cells for Accelerating Vaccines Development

Thesis Highlights:

• Adaptation of stable insect cell lines to hypothermic culture conditions allows to improve production of HIV Gag-VLPs (up to 26-fold) and Influenza VLPs (up to 12-fold);
• Adapted stable insect cells can reach 100x10⁶ cell/mL using perfusion, keeping specific Gag and HA production rates similar to batch, enabling to increase volumetric titer by 8‐fold;
• Adapted stable cells can be cultured at 20 x10⁶ cell/mL in continuous operation mode for 1.5 months, with specific protein (Gag and HA) production rates similar to batch - Gag-HA VLPs maintain their in size and morphology;
• The space time yield obtained in the continuous process using CSPR of 0.02 nL/cell.day outperforms batch and perfusion cultures.

Publications:

• B Fernandes et al (2022) Insect Cells for High-Yield Production of SARS-CoV-2 Spike Protein: Building a Virosome-Based COVID-19 Vaccine Candidate. MDPI Pharmaceutics 14(4), 854.
• B Fernandes et al (2022) Scalable process for high-yield production of PfCyRPA using insect cells for inclusion in a malaria virosome-based vaccine candidate. Frontiers in Bioengineering and Biotechnology
• B Fernandes et al (2021) Integrating high cell density cultures with adapted laboratory evolution for improved Gag‐HA virus‐like particles production in stable insect cell lines. Biotechnology and Bioengineering 118, 2536–2547.
• B Fernandes et al (2020) Adaptive laboratory evolution of stable insect cell lines for improved HIV-Gag VLPs production. J Biotechnology 307:139-147.
• J Vidigal et al (2017) RMCE-based insect cell platform to produce membrane proteins captured on HIV-1 Gag virus-like particles. Applied Microbiology and Biotechnology 102:655–666.

New Position: Senior Research Associate at Bayer Pharma Lab, at iBET.

Jorge Moura-Sampaio

PhD in Bioengineering, ITQB NOVA, NOVA University Lisbon (12^th May 2022)

Thesis Title: Structurally Primed Phage-display Libraries

Thesis Highlights:

• New NGS method for VL:VH pair analysis – results published in Computational and Structural Biotechnology Journal
• Improved affinity maturation success rates;
• New tool for antibody library comparison and automated candidate selection.

Publications:

• Jorge Moura-Sampaio et al (2022) Reconstruction of full antibody sequences in NGS datasets and accurate VL:VH coupling by cluster coordinate matching of non-overlapping reads. Computational and Structural Biology Journal 20:2723-2727.

New Position: Senior Researcher at the Novartis Pharma Lab, iBET.