Our Explorers share their experience

With Targenomix being focused on target identification of small molecules, the protein quality is a major concern for us. As we work with a wide range of different protein species, the only way to provide premium data is to routinely test our protein samples for their stability and quality.

nanoDSF is the method of choice for our quality control procedure which we now apply as the very first step in every project since it allows very quick measurements and provides detailed information on sample properties.

The broad concentration range of the Prometheus NT.48 allows us for the first time to investigate protein stability at very different protein concentrations. Since proteins can both be stabilized or destabilized with increasing protein concentrations, this analysis is crucial for the development of the experimental conditions for further binding assays, such as MST. Moreover, nanoDSF provides us key parameters for optimal handling and storage conditions. Related to this, we appreciate the high reproducibility and resolution of the unfolding data obtained with the Prometheus NT.48.

Dr. Sebastian Klie
CEO
Targenomix GmbH
Potsdam, Germany

We use this innovative technology to characterize advanced compounds, but also to confirm screening hits and to prioritize between chemical lead classes. We use the Monolith NT.115 from NanoTemper Technologies to determine binding affinities and expand its application range to thermodynamic characterizations…We observe a very good agreement of MST to other biophysical methods like surface plasmon resonance, X-ray structural information and molecular modeling.

Dr. Patrick Sieber
Lab Head R&D Cardiovascular and Fibrosis Biology
Actelion Pharmaceuticals
Allschwil, Switzerland

We have successfully implemented the Prometheus NT.48 device into our workflow for antibody characterization and optimization at Sandoz Biopharmaceuticals. We frequently use nanoDSF for thermal stability determination at different stages during the development of our biosimilar projects. It is an enabling technology for our projects since it reports with very high precision and high resolution on multiple unfolding transitions. We especially appreciate that the capillary-based measurement is not compromised by formation of protein aggregates since aggregates do not interfere with data collection. Intriguingly, the sensitivity and the resolution of the unfolding curves are not affected at all. Furthermore, we benefit from convenient usage, the instruments’ throughput and its little sample consumption when comparing to classical CD spectroscopy.

Dr. Alexander Bepperling
Lab Head Biophysical Characterization
Sandoz Biopharmaceuticals
Germany

At Novozymes we have implemented the nanoDSF technology for protein stability measurements. We are impressed by the ease of use of NanoTemper’s Prometheus NT.48 instrument particularly the low sample consumption and the wide concentration range it can measure under, which makes the Prometheus an ideal tool for our research. Not only does nanoDSF produce high quality data with excellent reproducibility, it also compares well to data we obtained with DSC, TSA and CD. All in all, the Prometheus NT.48 is a reliable research tool with a broad application range.

Dr. Werner W. Streicher
Senior Scientist
Novozymes A/S
Denmark

NanoTemper’s Prometheus is an excellent equipment for studying the Tm of any protein. It is comparable to DSC but is high throughput. Another advantage is that the analysis requires very small sample volume, negligible effort in sample preparation as even no dilution is required for concentrations of up to ~200 mg/mL in turn providing a broad concentration range for analysis. In addition to this, we can analyze 25 samples in less than 2 hours.

Sarita Sharma
Senior Research Scientist
Product Characterization Team
LUPIN LIMITED
Pune, India

nanoDSF technology is frequently used within the Biomolecular Interactions and Crystallization Core Facility of CEITEC. Most often we use Prometheus NT.48 to identify the optimal buffer conditions for protein stability prior to crystallization trials to improve the likelihood of protein crystals forming. Also, nanoDSF is used as a screening technique to test for the stabilizing effects induced by the binding of low molecular weight ligands or ions to the biomolecules. Prometheus NT.48 is a user-friendly instrument with little demands on sample consumption. Compared to standard DSF experimental setup, it is a label-free platform, which gives the data with very good precision. In my opinion, Prometheus NT.48 is an excellent choice for performing the stability assays of proteins.

Prof. Dr. Michaela Wimmerová
CEITEC
Brno, Czech Republic

We are using nanoDSF to determine refolding conditions on proteins purified from inclusion bodies. Before getting the Prometheus, this was more of a trial and error process. Thanks to Prometheus, we can increase the number of test conditions from a few to almost 50 using the same amount of test protein. Also, Prometheus saves us a considerable amount of time as we focus the later analytical SEC testing only on the protein samples from successful conditions.

Dr. Jacek Plewka
Post-doc
Jagiellonian University, Department of Chemistry
Kraków, Poland

We are very passionate in our quest to understand the molecular basis of the translation process. Our work focuses on the structural, biophysical and cellular outcomes of the functioning eukaryotic ribosome. In particular, we look at the GTPase associated center and its main functional element: the structure called P-stalk, which is thought to be responsible for the recruitment of translational GTPases and stimulation of factor-dependent GTP hydrolysis.
In the course of our study, the characterization of protein-protein interactions is of great interest, and we use a broad spectrum of biophysical techniques and methods to study the interplay of ribosomes with external factors. MST — the newest addition to our tool box — allows us to measure binding affinity with low sample consumption and the possibility of carrying out the measurements in solution without the need of immobilization

Dr. Przemyslaw Grela
Head of Protein Bioengineering Laboratory
Department of Molecular Biology
Maria Curie Sklodowska University
Lublin, Poland

The Prometheus NT.48 now allows us to quickly test the stability of proteins in different buffer conditions and even in the presence of detergents. This enables us to also measure membrane proteins in close to native conditions, which is not possible with the conventional DSF method. The unmatched resolution due to the high density of data points are very beneficial for antibody engineering projects since multiple transitions can be identified and determined with high accuracy and reproducibility. We also appreciate that the instrument is very robust and absolutely maintenance-free.

Dr. Thomas Schubert
CEO
2bind GmbH
Germany

The Monolith and the Prometheus instruments by NanoTemper Technologies have been used extensively by us at InStem for protein-ligand binding and protein stability measurements. The low sample volume requirements, easy instrument and software usage makes these devices popular among protein researchers on the campus of NCBS and InStem.

Dr. Vinod Nayak
Group Leader
InStem, NCBS Campus
Bangalore, India

I think nanoDSF technique is certainly cool and hassle-free. More important it requires very less sample and the instrument is maintenance free.

Dr. G. Bhanu Prakash Reddy
Scientist – Head of Biochemistry
National Institute of Nutrition
Hyderabad, India

The new Prometheus NT.48 is an extremely valuable tool in measuring thermal unfolding to quickly determine the melting temperatures of proteins or peptides, making life easy from using DSC and CD that consume more time and protein. The best part of the instrument is no cleaning business before the start of the experiment and the capillaries are affordable, also it is quick, precise and highly informative in reading the protein folding.

Dr. R. Nagender Rao
Project Leader (Dr. Sangita Mukopadhyay Team)
Centre for DNA Fingerprinting & Diagnostics
Hyderabad, India

The MST technology can be used as a complementary technology to the traditional methods in these fields. We can strongly recommend the MST technology for its low material consumption, short measurement times and broad application range in the field of molecule interaction studies. Using MST, we measured the Protein-Protein interaction that has been difficult to detect and quantify with ITC because of instability of the protein and very little heat changes of our samples.

Prof. Zhen Xi
Nankai University
Tianjin, China

The new 2nd Generation labeling kits from NanoTemper Technologies have improved the quantification of our measurements of intermolecular interactions and optimizing signal-to-noise ratios in our binding assays.  In general, the kits provide a quick and reliable option for labeling our proteins without any loss. The His-Tag labeling kit Red-tris-NTA 2nd Generation is one of the preferred consumables used by our visitors at the EMBL Biophysics Core Facility. We have seen optimal and reproducible binding results using this labeling strategy. In summary, we highly recommend the 2nd Generation dyes, which have helped us to execute several MST measurements in a short, and reliable matter using a very small amount of our purified proteins.

Dr. Maria Garcia
Lab Manager
European Molecular Biology Laboratory

Using MST and nanoDSF we could detect small molecule-protein interactions with high specificity, something that conventional biophysical techniques like SPR and intrinsic tryptophan fluorescence based studies had failed to detect. What gives nanoDSF and MST an edge over other techniques is their rapidity of analysis, ease of use and low material consumption. Also, NanoTemper Technologies’ Application Specialists provide great support, regarding all aspects from assay design, to sample preparation and final data analysis.

Shweta Karambelkar
Post Doctoral Fellow (Prof. Nagaraja Team)
Dept. of Microbiology and Cell Biology, Indian Institute of Science
Bangalore, India

We performed MicroScale Thermophoresis (MST) measurements to determine the dissociation constants of protease and substrate. We benefited from the low sample consumption, the small volumes and the possibility to choose the buffer freely especially to work at high salt concentrations. Furthermore MST allowed us to optimize assays in a straight forward manner and to carry out fast measurements resulting in highly reproducible data. The dissociation constants of USPs and various substrates measured by MST are in very good agreement with previously published affinities using alternative approaches like ITC, FA (fluorescence anisotropy) and SPR.

PD Dr. Günter Fritz
University of Freiburg
Freiburg, Germany

We use NanoTemper Technologies’ Microscale Thermophoresis to characterize the affinity of our compounds to their targets. Using this technology we obtained very convincing data that were in agreement with results obtained using other technologies like Surface Plasmon Resonance (SPR). We appreciated the relatively short time for the time to establish the assay compared to alternative methods to measure protein affinities as well as the comparatively low amounts of protein consumption. We conclude that this new technology gives rise to reliable quantitative affinity data.

Dr. Alexandra Matzke
CSO
amcure GmbH
Germany

Since many years we conduct research in the area of fast stability prediction methods for protein formulations. We have seen a continuous and still growing interest in the pharmaceutical community in such methods. With the Prometheus NT. 48 on board, our ability to contribute with precise and easy to obtain datasets was significantly improved. The new backscatter function makes the device particularly competitive.

Prof. Dr. Gerhard Winter
Ludwig-Maximilians-Universität München
Department of Pharmacy
Pharmaceutical Technology and Biopharmaceutics

A critical step in a protein’s purification process is to identify formulation buffers for optimal stability and activity. Before the Prometheus, we were primarily using a mixture of spectrophotometry, SDS-PAGE, size exclusion chromatography, multi-angle/dynamic light scattering, and various dyes used for aggregation and thermal shift assays. These methods gave us a stitched together picture of protein stability. However, each assay had its limitations. The more we manipulated the proteins to accommodate the compatibilities of the assays the less confidence we had that we were fully optimizing their stability. We were also sacrificing large amounts of our hard-fought purified material. Now with the Prometheus, we can get away with using 10uL of nanomolar concentrations of material per data point and acquire stability information all in one assay. We have been able to save on reagents, time, and costs with the throughput of this instrument. We have expanded the types and amounts of formulations we can test including denaturants, high salts, and detergents which are tricky for other methods. The Prometheus enabled us to identify stabilizing agents for complex proteins with multiple domains. The Prometheus technology is also being utilized for drug discovery efforts to identify small molecule binding to target domains. We continue challenging the instrument with complicated proteins and formulations; and it keeps giving us reproducible, high-quality data that allows us to get high quality therapeutics to patients faster.

Donnie Mackenzie
Scientist II
BioMarin Pharmaceutical

Tycho has become one of our standard methods for VLP analysis and evaluation. It uses low amounts of material, handling of samples is easy and provides a very fast estimation of VLP stability, concentration and purity.

Dr. Marcus Stapf
Senior Scientist
Neuway Pharma GmbH
Bonn, Germany

Cryo EM is a powerful method for gaining structural insights about molecules and their functional complexes in a solution. It is particularly important for understanding mechanisms of action of previously non-crystalizable protein targets. Such targets often prove to be difficult to handle and there is a strong need to verify the quality of sample entering CryoEM analysis. Tycho NT.6 is a great addition to our Cryo EM toolbox. It provides a fast and effortless way to analyze protein quality. It can be used at various stages preceding Cryo EM analysis to verify functionality of proteins – including protein purification steps, storage analysis or optimal buffer selection.

Dr. Sebastian Glatt
Head of the National Cryo-EM Facility, NSRC Solaris
Max Planck Research Group Leader, Malopolska Centre of Biotechnology
Deputy Director for Science, Malopolska Centre of Biotechnology
Jagiellonian University
Krakow, Poland

We use our Tycho for quick quality checks of our samples as well as for formulation analyses, and it helped developers to make changes to our manufacturing processes of recombinant vaccines. Tycho is also an easy-to-use tool to perform an accurate protein quantitation that is essential to our protein studies.

Julien Bouley, PhD.
Osivax

I’ve been using the Monolith NT.115 for almost 10 years and since then it has rapidly become a reliable, versatile and highly appreciated workhorse for my team projects. It uses single-use capillaries instead of chips or fixed sample holders and despite high usage and zero maintenance, our instrument never breaks down. You can test in almost unlimited sample types, including cell lysates without the risk of clogging or contamination. We have used Monolith to quantifying protein homodimerization in the presence of ligands or post-translational modifications, as well as to monitor slow interactions, which need minutes or hours to reach their equilibrium. The combination of ease of use, low sample consumption — a blessing for measuring low-affinity interactions — and user-friendly software —featuring a guided assay setup and quality checks — make it my team favorite analytical instrument. It is very important to thoroughly understand its principles and the characteristics of the chosen fluorescent label to obtain reliable results. However, in case of doubt or unusual results, the NanoTemper support team has always provided us with extraordinary assistance and advice.

Stefan T. Arold
Professor of Bioscience
King Abdullah University of Science and Technology (KAUST)

I am interested to determine affinities of a highly disordered protein derived from a coregulator to different nuclear receptor heterodimers. The MST method was very efficient to measure these affinities, with high reproducibility. Data are in perfect agreement with structural and functional data. I could not obtain such accurate data before with other methods, like fluorescence anisotropy. MST experiments consume very low amount of protein and are rapid to run, which is especially interesting when you want to do a number of runs in same conditions, but different interacting proteins. The support from NanoTemper Technologies in Brazil is very helpful and very competent. I will continue to use MST for other types of interactions (protein-peptide, protein-ligand, protein-protein).

Dr. Albane le Maire
LNBIO, CNPEM
Brazil

We are using MST for studying the activity of chromatin remodeling enzymes and to quantify their affinities towards DNA, RNA and other proteins. MST is a big step forward towards quantitative biochemistry. MST is simple to use and was quickly established in our lab. It is a good alternative to the traditional electrophoretic mobility shift assays.

Prof. Dr. Gernot Längst
University of Regensburg
Regensburg, Germany

At Celon Pharma we work with target proteins that lack enzymatic activity, which makes screening for potential drug candidates very challenging. MST provides us a new approach that brings a fast and reliable solution to this problem. With Monolith NT.115 we could finally perform single-dose screening with our target for which no other rapid screening methods were available. We expanded our use of Monolith to include it for the screening of small molecule-protein and protein-protein interactions in most of our ongoing R&D projects. The benefits offer by this technology over other methods are multiple: reduced costs due to low protein consumption, time saving for screening campaigns due to fast run times, easy setup for everyone in the lab to use, analysis software with useful tools and, last but not least, great technical support available 24/7. Summarizing, Monolith is a very valuable asset in our laboratory and should be considered by scientist for their screening campaigns.

Dr. Mikołaj Matłoka
Team Leader
Celon Pharma SA
Warsaw, Poland

We use MST to verify interactions between aptamers and proteins for drug development, and in the future, we plan to expand its use to study interactions between aptamers and small molecules. The reason why we were interested in MST in the first place was that we needed a method that required less amount of protein as they can be really expensive for us. Additionally, it provides very fast time to results, allowing us to analyze large batches of aptamers in relatively short time.

Dr. Anna Varizhuk
Senior Researcher
Biophysics Department
Research and Clinical Center of Physical-Chemical Medicine
Moscow, Russia

My main research at Sloan-Kettering Institute was based on biomolecular interactions between RNA-binding proteins and RNA and the discovery of small-molecules targeting that specific interaction. After optimizing and running FP-based high-throughput screen for small-molecules, the best and most useful technique to validate our hits was in fact MST with Monolith NT.115. Initially, we picked MST over other techniques for its versatility and high sensitivity, but later on we also benefited from the low sample size (protein and RNA), the easy set up and the very user-friendly analysis software. All of which meant we could achieve our research goals at a much faster pace. Remarkably, in few weeks we could perform optimal RNA-protein, RNA-small molecule and protein-small molecule interactions with very little assay development, labeling both nucleic acids and proteins with. Some of our MST data can be seen in Minuesa G. et al. Nat Commun. 2019, 19; 10(1):2691

Dr. Gerard Minuesa
Former Research Associate
Memorial Sloan-Kettering Cancer Center

The Prometheus nanoDSF enhances SAR for biologic engineering projects. We use Prometheus to evaluate molecules at early stages in HT-mode, taking advantage of its sensitivity to avoid costly scale up of candidates that will not move forward. Early screening by differential scanning calorimetry is not feasible with microgram-scale transfections or 1-day turnaround timelines that are routine in the early stages of antibody humanization. Thus, Prometheus enables the use of antibody domain stability as a critical ranking parameter in order to make the best choice of a stable antibody.
Additionally, the simultaneous acquisition of unfolding and aggregation data is more insightful than either alone. Because we could compare all aspects, Prometheus revealed an unanticipated interaction between domains of a non-antibody fusion protein that rendered the molecule unmanufacturable despite its robust activity in functional assays. Subsequently, we were able to better understand this molecule and re-focus engineering designs to address an unwanted molecular liability in our final designs.

Alan Kutach
Principal Scientist
NGM Biopharmaceuticals Inc.

My team’s goal is understanding the mechanisms and protein-protein interactions of the BCL-2 family of proteins in cell death and cell survival signaling pathways. We use MST for probing these molecular interactions and getting binding affinities because of the sensitivity of the method. This has benefited us particularly because our proteins are dynamic and prone to aggregation at the high concentrations required for other methods. With MST we can get affinities from proteins-protein and protein-small molecules interactions more reliably. We also use Tycho for routine check of the quality of our samples and MST assay development. Tycho is the perfect addition to the workflow because of its speed and label-free measurements.

Professor Evripidis Gavathiotis, Ph.D.
Departments of Biochemistry, Medicine and Experimental Therapeutics
Albert Einstein Cancer Center

MST allows us to easily explore the binding sites of plant enzymes and supplements classical kinetics measurements used in enzymology studies such as Kcat, Km or Ki, with the K_d. It also provides crucial information from site-directed mutagenesis experiments, when the key catalytic residues are targeted and the classical assays to determine kinetic parameters fail due to negligible or no activity of the mutants. With MST we can easily find out whether the mutation results only in loss of catalytic activity or binding affinity or potentially both.

Dr. David Kopecny
Assistant Professor
Department of Protein Biochemistry and Proteomics
Palacky University
Olomouc, Czechia.

MicroScale Thermophoresis (MST) is an in-solution, contact-free biophysical technology for protein-protein, protein-DNA and protein-small molecule interactions characterization. Label-Free MST allows to study biomolecules without disturbances by labeling or surfacing, and only requires little protein consumption and very short processing time. We are using this technology to determine the interactions between drug target protein and small molecules that have significant fluorescence interferences with other method and have obtained perfect results. We highly recommend MST for efficient and reliable quantification of biomolecule interactions in drug discovery.

Professor Liang Zhang
College of Basic Medical Sciences
Shanghai Jiao Tong University
Shanghai, China

NanoDSF (Prometheus NT.48) can analyze the stability of proteins in native conditions and get results quickly with extremely low sample demands. In addition, it has very high resolution and provides very detailed information from key unfolding transitions during intermediate processing. This generates reliable data for evaluating protein stability and makes our work a smooth and rapid process.

Hong Zhu
Department of Pharmaceutical Analysis
Chia Tai-Tianquing
Shanghai, China

The remarkable detection speed of Prometheus and the extremely low sample demands are very important for the screening of drug targets in the early development stage, especially for the screening of very important samples!

Xiao Luo
Vice President
Antibody Engineering Department
QiLu Pharmaceutical
Shanghai, China

In the past decade, my research has focused on characterizing multivalent binders against the spike proteins of the influenza A virus. MST made it possible to determine binding constants using whole virus particles, which revealed important insights on multivalent binders interacting with the native virus surface.

I highly recommend this valuable technology for virus binding studies.

Dr. Daniel Lauster
Freie Universität
Berlin, Germany

We initially rented Tycho NT.6 for a month and within this time, we were able to perform a large number of experiments that changed the course of our R&D dramatically. With the help of Tycho, we found out that our proteins behave differently from what we expected, and got a good idea of how to improve their quality. We have since been able to confirm our hypothesis with other methods and have made considerable progress in a very short time. I estimate that we would have needed at least a year to come to this level of knowledge using conventional methods such as gel chromatography and fluorescence assays. In light of this quick discovery, we were soon convinced to buy a Tycho for our R&D lab, and we are also considering to include it in our regular production processes.

Clemens Schneeweiß
Head of R&D Molecular Biology
imusyn
Hanover, Germany

In our group we apply protein engineering methods to optimize, for example, the activity and/or thermal stability of various biocatalysts. After rational or random mutagenesis, it is critical to verify the structural integrity and stability of the resulting enzyme variants. For this purpose, nanoDSF is really helpful as it enables an accurate and reliable measurement of melting temperatures of a large numbers of variants. We have also employed nanoDSF to test the influence of substrates, buffers, additives and cosolvents on protein stability of these biocatalysts. Importantly, the simple capillary filling procedure and intuitive software make the device easy to use, even for beginners.

Dr. Dominique Böttcher
Department of Biotechnology & Enzyme Catalysis
University of Greifswald
Greifswald, Germany

I would encourage everyone in my lab to use Tycho NT.6 as a check along all steps in a protein purification workflow. You might be able to better optimize a process you thought was already working by using the data from the NT.6.

Dr. Gregor Witte
Gene Center, Hopfner lab
Ludwig Maximilian University
Munich, Germany

The tremendous benefit of using nanoDSF (Prometheus NT.Plex) for the assessment of protein stability is the label-free, low sample volume and high-throughput capability which allows for screening various conditions in a short time and in a highly precise and easy-to-use fashion. Especially the close and fruitful collaboration with the NanoTemper team helps to unleash the full potential of the technology. Overall, nanoDSF has become a valuable key assay in our workflow.

Dr. Marius Müller
PostDoc and Subject Matter Expert
Janssen Switzerland

At our Proteomics Platform at La Trobe University (Melbourne, Australia), we have a Monolith NT.LabelFree and a Monolith NT.115. As an academic manager of the facility and group leader, I have been using both instruments for over 5 years. We have used MST to probe protein-small molecule and protein-protein interactions to guide our drug discovery program. One of the main advantages of using the MST is the low amount of samples required. I also frequently get told by people I train about how user-friendly the instrument and software are. The MST instruments have become an integral part of my research and our facility, and have led to several publications and collaborations. An important consideration for us is to minimize the amount of inhibitors used for initial screen, which we can do with the MST. We can also measure the interaction in solution, without the need for immobilization.

Dr. Tatiana Soares da Costa
ARC DECRA Fellow, Biochemistry
Department of Biochemistry and Genetics
La Trobe Institute for Molecular Science (LIMS)
La Trobe University, Melbourne, Australia

The simultaneous detection of micro- and macro-aggregates in combination with nanoDSF thermal unfolding profiles obtained with Prometheus Panta allow for very detailed characterization of biologics and protein-protein complexes. Our drug discovery and development customers  will benefit from this innovative instrument.

Dr. Thomas Schubert
CEO
2bind GmbH
Germany

We use Tycho across different groups in Protein Production, Biochemical Screening and Assay Development, and Crystallography – there are so many different uses for it, and new users need practically no training to get started.

Edoardo Fabini, PhD
Bioassay lab
Nerviano Medical Sciences
Milan, Italy

In any biophysical or biochemical experiment, it’s important to make sure the samples used are suitable for the job. We daily perform experiments on Tycho NT.6 to avoid poor to bad sample quality and to become more productive. We make use of data collected by Tycho in almost ten different projects to either analyze protein stability, or to test ligand binding to proteins in a thermal shift assay.

Another instrument that is part of our platform at Canther is the Monolith NT.115 Pico. Using MST, we already successfully determined K_ds of thousands of ligands against various types of proteins and oligonucleotides. Measurements have been done using GFP fused proteins extracted from cell lysates or using the Monolith His-Tag Labeling Kit RED-tris-NTA for his-tagged proteins.

Watch the pre-recorded webinar on how Romain Magnez, Technical Manager at Canther, used MST for the binding affinity analysis of PD-1 and PD-L1 directly in cell lysate.

Dr. Xavier Thuru, Scientific Manager
Romain Magnez, Technical Manager
Dr. Isabelle Van Seuningen, Laboratory Director

Canther
Lille, France