Science Pool

The periodic activation and inactivation of various cardiac ion channels enables the regular and synchronous contraction of the heart. Therefore, any disruption of the cardiac ion channel signaling network can result in a change in membrane potential and in the shape of cardiac action potential (AP). As a result, the cardiac excitation-contraction coupling can be modified which may lead to arrhythmias and potential patient death. Local Extracellular Action Potential (LEAP) is a technique which enables non-invasive, label-free monitoring of cardiac action potential in a high-throughput real-time format. In our research, we used the Axion LEAP MEA assay to analyse the effect of various selective and non-selective ion channel inhibitors on cardiac AP by quantification of action potential morphology, repolarization irregularities, and arrhythmic risk factors such as triangulation.

 

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Drug-induced cardiotoxicity may result from a functional change in cardiac electrophysiology (acute alteration of the mechanical function of the myocardium) and/or from a structural change, resulting in damage to the cardiac tissue. In our research, the effects of 42 reference compounds in human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) were investigated in a combined risk assessment strategy. Functional cardiotoxicity was evaluated through kinetic monitoring of calcium transients (CaT), while structural morphology changes and gross cytotoxicity were assessed using high-content imaging (HCI) and cellular ATP measurements. In addition, whole genome high-throughput RNA-sequencing (ScreenSeq) was performed in matched-sister plates. Data were analysed to determine differentially expressed genes (DEGs) and any associated perturbed pathways.

 

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The tuberculosis lesion environment is highly complex. Mycobacterium tuberculosis (M.tb) reside in various niches and their metabolism and other characteristics, including drug tolerance depend on the specific environmental characteristics of those niches. So-called persister bacteria refer to those that are difficult to eradicate with drug treatment and that may contribute to the long durations of TB treatment required for cure. Many in vitro models have been developed in attempts to mimic these niches with the objective of determining potential anti-TB compound activity in models that reflect M.tb’s characteristics in vivo.
Foamy macrophages Foam-M are characterized by lipid body accumulation induced by Mtb infection and they may be an important niche for Mtb during infection.

In this poster we:

• We describe the development of a robust 96 well plate Foamy macrophages assay designed to mimic this niche, and suitable for medium-high throughput evaluation of compound activity during drug discovery.
• We describe TB compounds activity in the foamy macrophage assay compared to other TB current assays
• We show that infection of foamy macrophage does not trigger an anti-inflammatory response in host cells

Read our poster to learn more about our research!"

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Hepatitis B is a serious liver infection caused by the hepatitis B virus (HBV). It is still a major public health problem with 250 million chronically HBV-infected patients worldwide with none of the current treatments leading to a cure.
More effective treatments are needed to achieve HBV cure in a large proportion of patients.
We aim to develop a treatment, combining simultaneous stimulation of CD40 and IFN-I pathway, which leads to a strong anti-HBV effect with minimal inflammation.

In this poster we:

• Present the key results that demonstrated the potential of such combination to induce anti-HBV effects in vitro and in vivo
• Introduce the design of our new antibody-based therapeutic that combine the two modalities in one single molecule
• Demonstrate the efficacy of the molecule in HBV infection system in Primary Human Hepatocytes
• Show key results from our preclinical safety assessment in vitro and in vivo in Non-Human Primate

Read our poster to learn more about our research!

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Full poster title: Development of recombinase-based targeted integration systems for production of exogenous proteins using transposon-mediated landing pads

Summary of the poster:

• We demonstrated proof of principle of targeted integration systems in our CHO host cell line with consistent genome integration into expected landing pad sites
• Test cases using three antibody or antibody-fusion therapeutic molecules showed similar levels of productivity
• We also show preliminary data from ongoing work to build upon these targeted integration systems, which includes isolating a single-copy landing pad cell line and developing a CHO display platform

Read our poster to learn more about our research!

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In this poster, presented at SfN 2018, Sessolo et al. present 3Brain high-density multi electrode array (HD-MEA) as a system to monitor and characterize seizure-like activity in hippo-cortical slices induced by different compounds.

The high system resolution allows to monitor in detail the entire slice and through the software showing the activity map (in real-time) the sign of compounds' action is easily found.

The technology allows to acquire Local Field Potential (LFP), Multi Unit Activity (MUA) and Single-Unit Activity.

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This poster includes information about:

• Functional brain slice electrophysiology by HD-MEA platform
• Combined neuronal circuitry studies through functional brain tissue imaging

Initially presented at FENS 2018 by Ugolini et al., 3Brain high-density multi electrode array (HD-MEA) as a system for long lasting monitor and characterize spiking activity of hundreds Purkinje cells simultaneously by using different positive and negative Ca++-activated K+ channels. Responses can be evaluated though different analysis. It is a useful tool for compounds validation on cerebellar slices.

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Toxicokinetic evaluation is a regulatory and scientific requirement in the drug development process. To obtain plasma, blood is generally withdrawn by a conventional venous collection method. Microsampling is a less invasive sampling technique, which allows to reduce the stress correlated to the conventional blood sampling and to decrease the number of rodents for a preclinical study. The implementation of microsampling in particular, in non-human primate can reduce the stress and promote a positive interaction with technical staff which improves the overall well-being of the animal (refinement).

In this poster we summarise the work done to evaluate the possible influence of the blood sampling method on drug plasma concentrations, using LC-MS/MS methods in non-human primate for four drugs selected based on acid-base and volume of distribution properties.

The poster was presented by our expert Rossella Cardin at the 24th International Reid Bioanalytical Forum held in Cambridge, UK, on June 13-16, 2022.

 

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Key Takeaways:

• Non-infectious, purified RVLPs can be used in place of model viruses to predict process performance for viral clearance.
• Initial screenings show that clearance also trends similarly between bench screenings and higher throughput plate screenings.
• Plate-based screening of RVLPs in-process can examine up to 24 different run conditions simultaneously and uses less viral surrogate compared to bench scale runs, allowing for greater evaluation and confidence going into formal viral clearance studies.

Improving virus clearing studies in recombinant protein production

Chinese hamster ovary (CHO) cells are the most frequently used mammalian host cells for the industrial manufacturing of recombinant protein therapeutics. They can produce recombinant proteins on the scale of up to 10 gram per liter of culture. However, they are also known to contain type‐C endogenous retrovirus (ERV) sequences in their genome and to release retroviral‐like particles. Although evidence for their infectivity is missing, this has raised safety concerns, and regulatory agencies require demonstration that the purification process removes or inactivates viruses.

Viral clearance validation is assessed through “spiking studies”, whereby model mammalian viruses are introduced into process material which then undergoes the purification technique to be tested. Viral quantity before and after processing is determined through infectivity or qPCR assay. As these studies use live viruses, they require specialized Biological Safety Level laboratories (BSL) and experienced personnel and can create a substantial bottleneck because typically only 3rd party facilities are qualified to perform these studies.

As an alternative, Just - Evotec Biologics is in the early stages of establishing a high-throughput process using commercially available purified retrovirus-like particles from Cygnus Technologies LLC. These particles are non-infectious and mimic the physicochemical properties of live infectious viruses. By using these particles as spiking agents, the retroviral clearance capability of downstream unit operations can be studied, assessed, and quantified by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR). Usually, this is performed at bench scale using chromatography columns.

In a poster presented at this year’s ACS spring conference entitled High throughput optimization of chromatography steps for viral clearance using retrovirus-like particles (RVLPs), researchers from Just-Evotec Biologics detailed the high-throughput workflow for the analysis of RVLP content for rapid analysis of in-process samples.

The research team compared common bench scale chromatography runs with a plate-based screen using resin-loaded filter plates and a liquid handling robot. While at bench scale, only a single set of run conditions can be tested at a time, the plate-based screening can examine up to 24 different run conditions simultaneously. It also uses less RVLP stock solution. The researchers expect that plate-based screening of RVLPs will not only save time and costs, but also allows for better evaluation and confidence before formal viral clearance studies.

 

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Autotaxin (ATX) is a secreted glycoprotein that hydrolyzes lysophosphatidylcholine (LPC) to lysophosphatidic acid (LPA). LPA signalling directly modulates tumour cell function and contributes to the development of the fibrotic tumour microenvironment, resulting in reduced host immunity and impaired response to therapy.

iOnctura, a clinical-stage oncology company, based in Switzerland (Genève), targeting core resistance and relapse mechanisms at the tumor-stroma-immune interface, has developed a potent and orally available autotaxin inhibitor, IOA-289, as a novel treatment for pancreatic cancer and other highly fibrotic tumours.

The first Phase I clinical study in healthy volunteers has been successfully completed. Evotec is very proud to have actively contributed in conducting the trial and analyzing the results that led to the achievement of this exciting goal.

Read the poster presented by iOnctura at the ESMO Immuno-Oncology Congress held in December 2021 to learn more about IOA-289 and its activity as autotaxin inhibitor. IOA-289 was also presented by iOnctura at the recently held AACR Annual Meeting in New Orleans.

 

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