Dr. Thomas Machleidt leads the Advanced Technology Group within Promega’s Research & Development organization, where he focuses on the development of innovative analytical tools for life science research. His work centers on engineered reporter proteins, detection chemistries, and novel assay formats that enable deeper insights into cellular processes.

Dr. Machleidt earned his M.S. in Biology from Eberhard Karls University in Tübingen and completed his Ph.D. in Immunology at the Technical University of Munich. He went on to hold research and development positions at Johnson & Johnson, Ansata Therapeutics and Life Technologies. Over the course of his career, Dr. Machleidt has authored or co-authored more than 70 peer-reviewed publications and is listed as an inventor on over 20 patents.

 Innovation in Bioluminescent Technologies -  Analyzing Endogenous Protein Dynamics

In this presentation we will introduce some of the latest advanced technologies and workflows Promega has developed for analyzing endogenous protein dynamics using luminescence and fluorescence. DualTag is a compact (33-aa) tag that combines luminescent and fluorescent detection, enabling robust assay performance and compatibility with imaging and flow cytometry. A second technology based on a chimeric proteins combining circularly permuted NanoLuc with HaloTag enables robust dual-color luminescence detection in imaging and plate based assays. This supports multiplexed assays to monitor target and control proteins simultaneously. Lastly, CRISPR-based workflows were used to create endogenous protein-protein interaction models (e.g., EGFR/GRB2, RAS/RAF) using NanoBRET™ and NanoBiT® reporters. These models preserve native signaling, support high-throughput screening, and have been applied to 3D cultures and isogenic variants to study drug resistance

Originating from biosciences, I moved to the Faculty of Chemistry to introduce methods for testing the bioactivity of experimental molecules in the field of medicinal chemistry. Currently, I’m a leader of the Bioorganic and Medicinal Chemistry Group. As a researcher, I'm involved in developing small molecules intended to block such protein-protein interactions as PD-1/PD-L1 or MDM2/p53, especially including new binding modalities and mechanisms of action. I'm also devoted to developing new cell-based models, including co-culture models, for testing bioactive compounds.

Testing of PD-1/PD-L1-targeting molecules with cell-based models

Targeting immune checkpoint molecules (ICMs) with therapeutic monoclonal antibodies (mAbs) is an established anticancer treatment strategy. Reliable in vitro models are required for discovering new, non-mAb immune checkpoint inhibitors (ICI), such as small molecules or peptides. Cell-based immune checkpoint blockade assays provide such models, as exemplified here for the compounds targeting the PD-1/PD-L1 interaction. The analysis of resulting data allows not only for defining and comparing the bioactivity of the tested ICIs but also for estimating their nonspecific toxicity. Additionally, an alternative assay setup provides information on the response of primary cells to the treatment. The talk will provide a brief view of the progress in PD-1/PD-L1-targeting ICIs developed in our laboratory.

Wojciech Wiertelak is an assistant professor in the Department of Biochemistry at the Faculty of Biotechnology, University of Wrocław, where he actively engages in both teaching and research. He obtained his PhD in biological sciences in 2024, and his primary research interests revolve around glycosylation, with particular emphasis on congenital disorders of glycosylation. He focuses on studying glycosylation complexes that involve nucleotide sugar transporters and glycosyltransferases, employing bioluminescent techniques to investigate protein–protein interactions.

Analysis of Golgi apparatus protein complexes involved in glycosylation using the NanoBiT technique

Glycosylation is one of the most important post-translational modifications in eukaryotic cells. It involves the covalent attachment of sugar residues primarily to proteins or lipids. This multi-step process requires the coordinated action of numerous enzymes. Nucleotide sugars, which serve as substrates for glycosylation reactions, are synthesized in the cytosol and subsequently transported into the Golgi apparatus or the endoplasmic reticulum via specialized nucleotide sugar transporters. Once inside these organelles, they are utilized by glycosyltransferases to modify target molecules. Protein–protein interactions constitute one of the regulatory mechanisms overseeing this process, influencing everything from enzyme localization to overall pathway efficiency. The presented findings highlight the potential of the NanoBiT technique, a sensitive luciferase-based assay, for examining interactions between Golgi-resident membrane proteins. By revealing the dynamics of these complexes, this approach provides valuable insights that could advance our understanding of glycosylation-related disorders and inform the development of novel therapeutic strategies.

Dr. Stephan Kirchmaier supports scientists across Europe in the pharmaceutical, biotech and academic sectors by connecting them with innovations from Promega’s R&D. He studied molecular biology in Vienna and at the University of Groningen, and earned his PhD from Heidelberg University. Before joining Promega in 2021, he conducted postgraduate research at the German Cancer Research Center.

Advancing Cellular Analysis with Bioluminescence Imaging

Bioluminescence imaging offers a valuable alternative to fluorescence-based methods, particularly for long-term and dynamic studies of endogenous biological processes.
Unlike fluorescence microscopy, it avoids issues such as phototoxicity and photobleaching, making it suitable for sensitive cell systems and extended observations. This talk presents a bioluminescence imaging approach using the GloMax® Galaxy Bioluminescence Imager to visualize dynamic events, track the localization of molecules, and assess cellular morphology. The method supports both live-cell and fixed-cell imaging and allows integration of assay-specific signals with cellular reference markers. We will discuss applications ranging from basic cell biology to long-term imaging experiments supported by environmental control systems.

Dr. Wnorowski is a molecular pharmacologist serving as a Principal Investigator at Biotechna SA. He is focused on developing novel compounds with receptor-mediated and anti-cancer activities. He holds PhD from the Medical University of Lublin, where he is an Associate Professor. Dr. Wnorowski has led or participated in drug discovery projects funded by major Polish agencies, including FNP, ABM, NCN, and gained international experience at National Institute on Aging (NIA/NIH, USA), Saarland University (Germany), and Vlaams Instituut voor Biotechnologie (VIB, Belgium). He has authored over 60 publications and holds 3 patents.

Avoiding Misleading Data: The Power of Combining Multiplex Assays with Dose & Time Response

Interpretation of in vitro cell-based pharmacology data is critical, yet reliance on still common single-dose, single-time point, single-assay experiments can be highly misleading, potentially masking complex drug behaviors or suggesting incorrect mechanisms. This presentation argues for the power of integrating comprehensive dose-response analysis, time-kinetics, and multiplex assays to generate more reliable and informative data. It will be explored how kinetic dose-response studies reveal crucial dynamics shifts over time. Critically, the benefits of multiplexing will be highlighted, demonstrating how simultaneously measuring multiple endpoints (e.g., metabolic activity, cell number, and membrane integrity) within the same well provides richer, context-dependent insights and internal validation. Examples will be presented contrasting misleading conclusions drawn from single-point data versus the nuanced understanding gained from the integrated, 'all-at-once' approach. Attendees will learn how this strategy enhances data interpretation, avoids common pitfalls, and ultimately yields a more accurate picture of drug action in cellular systems.

Coming soon

Bioluminescent Monitoring of Inflammation in Neurovascular Unit (NVU) Model for Drug Testing in Depression

In our pursuit to develop a sophisticated three-dimensional (3D) neurovascular unit (NVU) model using Brain-on-Chip technology for drug testing in depression, we have integrated Promega's bioluminescent assays to enhance the accuracy and efficiency of our evaluations. Specifically, the CellTiter-Glo® Cell Viability Assay has been instrumental in assessing cell viability within our 3D cultures, as well as determining optimal hormone concentrations for inducing inflammatory states within the NVU model.

Currently, our aim is to effectively induce an inflammatory state within our NVU model utilizing hormones (cortisol, angiotensin, aldosterone) and inflammatory cytokines (IL-1β and TNF-α). Initially, the inflammatory response was validated using Promega’s RealTime-Glo™ MT Cell Viability Assay in 96-well plates, allowing real-time monitoring of cell metabolic activity under inflammatory conditions. Subsequent evaluations employed the Lumit® IL-1β Human and Lumit® IL-8 assays to precisely quantify cytokine levels, providing clear insights into inflammatory dynamics within the NVU model. Furthermore, the Caspase-Glo® 1 Inflammasome Assay was utilized to detect inflammasome activation, critical for understanding inflammation-related pathology in depression. By incorporating these advanced bioluminescent tools from Promega, we achieved a comprehensive understanding of inflammatory mechanisms and cellular interactions within the NVU, establishing a robust platform for drug permeability studies, mechanistic insights, and novel therapeutic testing for depression and related neurological disorders.

I am a researcher at the Faculty of Medicine and Dentistry, Palacký University Olomouc, specializing in molecular biology and experimental medicine. My work focuses on developing high-throughput screening tools and disease models to support drug discovery for chronic and rare diseases. I have expertise in CRISPR/Cas9, fluorescence-based assays, tissue cultures, and high-content data analysis. My research integrates molecular tools and imaging technologies to improve screening workflows and enhance translational relevance in preclinical settings.

Illuminating the path to precision therapeutics in cystic fibrosis

Cystic fibrosis (CF) is a genetic disease caused by mutations in the CFTR gene, affecting protein biosynthesis, trafficking, or function. While common mutations are well-studied, rare CFTR variants remain underexplored. To address this, we developed a novel bioluminescent assay using a NanoBiT-based approach. By tagging endogenous CFTR in a physiologically relevant context, this model enables live-cell detection of CFTR at the plasma membrane. It provides a robust platform for introducing and characterizing rare CFTR mutations, as well as evaluating their functional rescue by candidate therapeutics. This assay offers a valuable tool for understanding rare CF variants and accelerating personalized treatment strategies in cystic fibrosis.

Małgorzata Adamiec-Organiściok, PhD is a research assistant at the Department of Systems Engineering and Biology, Faculty of Automatic Control, Electronics and Computer Science, Silesian University of Technology in Gliwice, Poland. In 2023, she obtained her doctoral degree in biomedical engineering.

Her research focuses on cell biology, particularly the mechanisms of programmed and non-programmed cell death, including ferroptosis. She investigates the role of oxidative stress in regulating cell death processes across various cell lines, aiming to better understand cellular responses to external stressors such as ionizing radiation.

Ferroptosis: A Form of Cell Death Driven by Oxidative Stress

Ferroptosis is an iron-dependent form of programmed cell death closely associated with oxidative stress and redox imbalance. Key roles in this process are played by reactive oxygen species (ROS), lactate dehydrogenase (LDH), and glutathione (GSH), the primary intracellular antioxidant. Different cell lines exhibit varying sensitivity to ferroptosis induction, potentially due to differences in LDH activity, baseline ROS levels, and GSH availability. Elevated ROS levels and GSH depletion promote lipid peroxidation, ultimately leading to cell death, while LDH activity can reflect the metabolic state and the extent of cellular damage. In this study, bioluminescence-based assays were used to quantitatively assess ROS, GSH, and LDH levels, enabling sensitive and real-time monitoring of metabolic changes. Due to its high sensitivity and low background interference, bioluminescence provides a precise tool for detecting differences in oxidative stress responses among various cell lines during ferroptosis induction. The results contribute to a better understanding of cellular resistance mechanisms, particularly in cancer cells, and support the development of redox-targeted therapies.

I hold a PhD in cell biology from the University of Warsaw. Since 2019, I have been working at Adamed Pharma S.A., where I am part of the Drug Discovery Department in the Cell Biology and In Vivo Research Team. My scientific work focuses on in vitro studies of biologic drugs with therapeutic potential. My core expertise includes techniques such as droplet digital PCR (ddPCR), flow cytometry, spectral cell sorting, and confocal microscopy.

Development of novel incretin agonists using luciferase, fluorescence, and NanoBRET in vitro assays in the DILOC-2 project

Incretin hormones—GLP-1, GIP, and glucagon—play essential roles in glucose homeostasis and energy metabolism by activating GLP1R, GIPR, and GCGR, respectively. These pathways are key pharmacological targets in obesity and type 2 diabetes treatment. The DILOC-2 project began when therapeutics targeted only GLP1R, initially aiming to design a multiagonist peptide acting on GLP-1, GIP, and GCG receptors with efficacy comparable to existing GLP1R-based drugs. Later efforts focused on surpassing current clinical candidates like retatrutide and approved agents such as tirzepatide. The project involved extensive development and refinement of analytical methods and bioassays to evaluate compound activity and therapeutic potential. During the presentation, I will provide an overview of the in vitro studies utilized cAMP-based assays for human GLP1R, GCGR, and GIPR to measure intracellular cAMP response to peptide binding. Additionally, I will discuss NanoBRET assay for receptor-ligand interactions at the cell surface.