In the first article in this series, we looked at how HMGB1 has taken an increasingly important position as a key mediator in the immune response, playing a major role in many diseases, from cancer to coronavirus. There is now significant evidence that HMGB1 is essential for SARS-COV-2 replication, as well as potentially being a therapeutic target in severe cases of COVID-19.1 In this article we examine how we can effectively measure HMGB1 accurately in serum and other samples and begin the journey from research to clinic.
How the human body deals with infection depends on an individual’s immune response. When looking at the body’s response to SARS-CoV-2, the state of the immune system has a crucial impact on the clinical outcome. For example, HMGB1 (High Mobility Group Box 1) protein is a key mediator of the immune system, and as such it has been shown to be critical in the replication of SARS-CoV-2. This article outlines the potential roles of HMGB1 in the race to find solutions to the coronavirus pandemic.
In the first article in this series, we looked at how HMGB1 has taken an increasingly important position as a key mediator in the immune response and as such plays a major role in a large number of diseases – from sepsis to cancer. As Professor Helena Erlandsson Harris, a pioneer in HMGB1 research, says, “I am convinced that the next step will be even better data to demonstrate the usefulness of HMGB1 as a prognostic/diagnostic biomarker. This has been hampered by the need to understand the isoforms that control different functions and also the methods for measuring HMGB1. It would be even better if HMGB1 detection were included in larger biomarker panels.” HMGB1 has indeed been included as a necessary biomarker in consensus guidelines for the detection of immunogenic cell death. The question is how to measure it. In this article, we will look at the development of increasingly sensitive, reliable and easy-to-use assays for clinical research and routine use and how this has been complicated by the need to resolve the isoforms, and also overcome interference caused by auto-antibodies and other proteins that naturally interact with HMGB1 to modulate its function.
As a nuclear protein present in most cell types, HMGB1 (high mobility group box 1) is a key mediator of the immune system in health and disease. Interest in HMGB1 has increased dramatically as the protein has been shown to be critical to the cell’s response to stress and plays a major role in many disease states, including infectious diseases, ischemia, immune disorders, neurodegenerative diseases, metabolic disorders, and, not least, cancer. Highly conserved in mammals, HMGB1 (also known as HMG-1 and amphoterin) is primarily located in the chromatin where it stabilizes chromosome structure and plays a key role in controlling gene expression.
Last night you were up until midnight tending to your live-cell experiment. This morning you woke up with great expectations, only to find that your cells are sick and the entire experiment must be repeated. Sound familiar? It happens all too often, and the consequences can be heartbreaking – deadlines missed, expensive reagents wasted, precious samples lost.
Cell-based assays are a core research tool, offering an informative and cost-effective counterpart to in vitro and animal tests. Where destructive methods involving cell lysis once predominated, live cell assays are now commonplace, with measurements collected in real time, either at a single time point (end-point assays) or repeatedly over the course of minutes, hours or even days (kinetic assays).
Imagine life science research without cell-based assays. Or without cultured cells of all types to power those assays. Healthy, high-quality cells at the right point of confluence are vital for proliferation, kinetics, cytotoxicity, and gene expression studies particularly during long-term experiments. With so many different cell types, assay formats, and detection methods the variability inherent in cell-based assays can be enormous. There’s no room for inconsistency in cell counts and confluence assessments — it’s counterproductive and just wastes time. What’s the best way to improve counting accuracy in your cell-based assays?
You may be convinced that your academic research laboratory is humming along just fine and cannot benefit from, take the time to consider, and perhaps most of all, afford adding automation to your workflow.
Achieving reproducibility in a cell-based fluorescence assay can be a real challenge. For example, you might run a very basic experiment to determine the optimal concentration of cell media supplements for your primary cell line. You're looking for differences in proliferation.
How can we improve upon the completely artificial situation that we have today for screening drugs? We spoke to Dr. Christopher Millan, Co-Founder and CTO of the up-and-coming company, CellSpring. Based in Zürich, Christopher Millan with his business partner, CEO Kramer Schmidt, are both Americans. We also asked Chris how two Americans end up establishing a biotech start-up company in Switzerland.
With today's demands of throughput and flexibility, how can you perform screening better? We spoke to Dr. Bernhard Ellinger, Principal Scientist at the Fraunhofer Institute for Molecular Biology and Applied Ecology. Dr. Ellinger is one of the first testers of the Fluent®* 780 liquid handling automation platform from Tecan. He put the system through its paces in a diverse range of applications for over 3 years. Here’s what he learnt.
With multiple tests to perform on a tiny volume, samples are getting more precious. And as Next Generation Sequencing pushes the envelope on cost and throughput, scientists are looking for ways of reducing reagent volumes without compromising on quality. Tecan has a tip.
“When you can measure what you are speaking about, and express it in numbers, you know something about it.” Lord Kelvin knew that. To be confident in your results, to quickly move your studies forward, and to be the first to publish your conclusions, you need to know that your numbers are right. The proof you need lies in reproducibility, and reproducibility in any cell-based assay starts with accurate cell counts.
When it comes to drug development, the challenge is always to create as much in-vivo relevant data as possible. The more relevant in-vivo data you can gather, the lower the risk of the drug not passing a clinical trial.
What are the benefits of the new Spark® 20M when it comes to accelerating the drug discovery process? This presentation from SLAS2016 goes beyond discussing typical microplate readers and washers to covering processes for optimizing assay development.