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Extracellular vesicles (EVs) are membrane vesicles fundamental for cellular communication processes. Either by taking advantage of EVs innate properties, or by loading them with therapeutic agents, EVs have emerged as cell-free strategies with diverse clinical applications. Indeed, when compared to mesenchymal stromal cells (MSCs), similar or better therapeutic benefits have been reported for MSC-derived EVs, thus explaining the increased interest in EVs over the years. Despite appealing features, EVs implementation in the clinic is still limited by technical difficulties, like robust isolation methods capable of delivering a high-quality product with high yield. In this work, published in the journal Separation and Purification Technology, we described anion exchange chromatography as a robust and better alternative to already implemented techniques. Following this strategy, more than 85% of injected EVs could be recovered, while complying with the stringencies set by regulatory agencies. This study developed at iBB was led by Dr. Ana Fernandes-Platzgummer and Prof. Ana Azevedo and co-first-authored by PhD students Ricardo Silva and Sara Sousa Rosa.

In recent years, there has been an increased interest in exploring the potential of micro-and mesoscale milling technologies for developing cost-effective microfluidic systems with high design flexibility and a rapid microfabrication process that does not require a cleanroom. Nevertheless, the number of current studies aiming to fully understand and establish the benefits of this technique in developing high-quality microsystems with simple integrability is still limited. In a recent paper published in Micromachines, BERG-iBB researches, Ana Azevedo and Amin Javidanbardan, in collaboration, with João Pedro Conde and Virginia Chu from INESC-MN, describe a systematic methodology for developing 3D high-quality thermoplastic-based molds for microfluidic structures based on micromilling technology. The work was performed under the Marie Skłodowska-Curie grant agreement No. 812909 CODOBIO, within the Marie Skłodowska-Curie European Training Networks framework.

Vaccines are one of the most important tools in public health and play an important role in infectious diseases control. mRNA vaccines are reaching the stoplight as a new alternative to conventional vaccines due to their precision, safe profile and flexible manufacturing. In fact, the first Covid-19 vaccines to receive regulatory approval are based on mRNA technology. In a joint review just published in Vaccine, Sara Sousa Rosa, Miguel Prazeres, Ana Azevedo from BERG-iBB, together with Marco Marques from University College London, describe the current state-of-art of mRNA vaccines, focusing on the challenges and bottlenecks of manufacturing that need to be addressed to turn this new vaccination technology into an effective, fast and cost-effective response to emerging health crises.

Ana Azevedo from BERG-iBB was invited to deliver a presention on continuous purification of antibodies at the European Summit of Industrial Biotecnology, in a special session dedicated to the Continuous Downstream Processing of the Biomolecules of the future. The section included also presentations from Novartis, Merck and Sanofi. The summit was held on the 18th – 20th November 2019, in Graz, Austria.

The mechanisms underlying the adsorption of monoclonal antibodies (mAbs) towards phenylboronate (PB) ligands have been elucidated by Flow Microcalorimetry. The knowledge acquired BERG-iBB researchers, in collaboration with Cristina Dias-Cabral's group from CICS-UBI, suggest how to increase the salt tolerance of the bond between ligand and biomolecule by manipulating the environmental conditions.It was demonstrated that such feature can be accomplished either by using mobile phase modulators that promote the specific cis-diol interactions between PB ligands and mAbs or by diminishing the non-specific interactions involved in the adsorptive process. The research team believes that these findings could contribute towards the effective integration of boronate chromatography in the current mAb downstream processing platform. The paper was published in Biotechnology Journal.

The conference "Integrated Continuous Biomanufacturing IV", organized by ECI (Engineering Conferences International), was held in Cape Cod, Massachusetts, in October 6-9. Ana Azevedo from BERG-iBB chaired session 5 on the "Validation of Integrated Continuous Processes" together with Raquel Orozco from Boheringer Ingelheim. Ana was further selected by the steering committee to chair the next ICB conference in 2021.

The establishment of novel, simpler and improved purification processes is extremely important to make antibody-based treatment options more accessible and with a lower economic burden to healthcare systems and patients. In this context, the development of microfluidic tools for the optimization of chromatography operating conditions can offer advantages such as low consumption of molecules, fast generation of results and ability to automate and multiplex the assays. In a collaboration between INESC-MN and iBB, researchers have recently reported a novel microfluidics-based methodology to perform rapid and multiplexed screening of various chromatography ligands relative to their ability to bind different target molecules in solution. The work was published in the Biotechnology Journal and is expected to contribute to the fields of separation sciences and analytical chemistry and inspire new developments within the purification of biopharmaceuticals or other molecules.

Rapid empirical optimization/screening of the partition of biomolecules in aqueous two-phase systems (ATPS) can be performed in microfluidic devices coupled with fluorescence microscopy. However, the effect of fluorescent tags on partition must be carefully evaluated to assess the validity of extrapolating the results to a situation where the molecules are unlabelled. To evaluate this influence, researchers from BERG-iBB and INESC-MN (Mariana São Pedro, Ana Azevedo, Maria Raquel Aires-Barros and Ruben Soares) carried out an ATPS study using both microtubes (batch) and microfluidic devices (continuous). IgG molecules were labelled with three fluorescent tags with different global charge and reactivity, namely BODIPY™ FL maleimide, Alexa Fluor® 430 NHS ester and BODIPY™ FL NHS ester. The thiol reactive BODIPY™ FL maleimide coupled with a reducing agent provided minimal bias relative to the native molecule in PEG-phosphate systems. The work was recently published in the Biotechnology Journal.

Description: A plaque used to determine bacteriophage titers, featured photo by Jorge João, Copyright BERG-iBB 2018.

Context: BERG researchers led by Ana Azevedo and Miguel Prazeres are developing processes for bacteriophage manufacturing in collaboration with the portuguese start-up Tecnophage. The work is supported by the Portugal2020 project "Phage4BacID-Bacteriophages in the Diagnostic and Prevention of Multiresistant Infection in Portugal and in the World".

The development of new scale-down models for accelerated and cost-effective biopharmaceutical process development has been receiving considerable attention in the last decade. Microfluidic systems have emerged as a particularly attractive candidate by offering miniaturization, parallelization, and allowing continuous and precise pumping and online monitoring possible. In a collaborative work, published in Journal of Chromatography A, BERG-iBB researchers Ana Azevedo and Amin Javidanbardan, together with INESC-MN researchers João Pedro Conde and Virginia Chu, propose a simple and cost-effective method for developing microfluidic chromatographic systems with integrated sensors. Fabrication and integration of microsensors, in this case, an impedance sensor, in each microcolumn provided the possibility of parallelizing experiments with separate online monitoring. The work was performed under the Marie Skłodowska-Curie grant agreement No. 812909 CODOBIO, within the Marie Skłodowska-Curie European Training Networks framework.

Amin Javidanbardan won the Eppendorf Poster Award at the 6th BioProScale Symposium “Innovative scale-up and scale-down for bioprocess intensification.” The event took place virtually from 29.03. to 31.03.2021. Amin presented his poster with the title “Rapid and cost-effective fabrication of microchromatography integrated with microelectrode impedance sensor.” Amin, a Marie Curie Early-Stage Researcher of the ITN action “CODOBIO,” is enrolled in the Ph.D. program of Biotechnology and Biosciences and is being supervised by Ana Azevedo (iBB-BERG) and João Pedro Conde (INESC-MN), both from the Department of Bioengineering. Congratulations Amin!

Aqueous two-phase extraction (ATPE) can bring numerous benefits to the downstream processing of biological products, including high recovery yields, high selectivity and relatively low cost. With biomanufacturing entering an era where continuous processing is regarded as the key solution to increase flexibility and decrease costs, the intrinsic characteristics of ATPE and high degree of integration, makes it highly suitable for continuous processing. In a recent paper published in Fluid Phase Equilibria, Diogo Faria and colleagues from BERG-IBB make a brief review of the history of continuous ATPE, analyse the main devices used in continuous extraction, and discuss the key and future challenges in the development of continuous processes.

Diogo Faria, PhD candidate at BIOTECnico Programme in IBB-BERG, gave a poster presentation at the conference "Integrated Continuous Biomanufacturing IV", organized by ECI (Engineering Conferences International), held in Cape Cod, Massachusetts, in October 6-9. The poster was about Continuous Aqueous Two-Phase System Extraction using Oscillatory Flow Reactor. The work was carried out under the supervision of Prof. Ana Azevedo (IBB-BERG) and Prof. Raquel Aires Barros (iBB-BERG), with colaboration of António Ferreira (FEUP).

The first consortium meeting of the EU-funded ITN CODOBIO: Continuous Downstream Processing of Biologics was held on the 22nd October at IST, Lisbon. The consortium involves 20 participants from Academia, Industry and Regulatory Agencies. Thirty two members were present in the meeting including the 15 Early Stage Researchers. Amin Javidanbardan and Malik Wahab from BERG-iBB had the opportunity to present their PhD projects, which focus on the miniaturization of continuous bioprocesses using microfluidics.

The research performed by Ana Azevedo and Carla Carvalho from BERG-iBB was recognized in this year's edition of the ULisboa/CGD Scientific prizes. Ana won the first prize Ex Aequo and Carla received and honorable mention for their work in the category "Biology, Biological Engineering, Biochemistry and Biotechnology". The award ceremony took place on the 8th April.