See Springer Lab get Explorer Award for looking at protein stability of MHC class I molecules
Read what our other customers have to say about NanoTemper
Leading the biophysics team in JBIO at Janssen since 2017, my goal is to build an industry leading biophysics team using all high throughput screening assays to enable rapid lead selection. Two of the critical properties of mAbs are conformational and colloidal stability. These play an uncompromised role in determining both in-vivo behavior and successful manufacturability. The first instrument that I bought to build out our HTP screening for hit selection is the Prometheus. While there are other HTP instruments for determining the thermal stability of molecules, they were not up to my expectations in terms of quality and reliability of data, material consumption, analyst friendliness, and time required to generate the data.
Prometheus nanoDSF has become part of our routine HTP screening workflow that uses only micrograms of purified proteins. We have done 1000s of experiments using Prometheus to determine the intrinsic stability of hits in different experimental conditions, and Prometheus continues to provide the quality and reliability of Tm and Tagg determination our work demands. Our data has also supported the accuracy in the measurement of Tm as verified with other methods such as DSC. I am very pleased with this discovery and highly recommend Prometheus to be part of antibody discovery and development workflows at any company.
Read how Prometheus helped Dr. Venkataramani’s stability enhancement study
Sathya Venkataramani, PhD
Associate Director, Intrinsic Property Assessment – Biophysics Group
Biologics Development Sciences Department
Janssen Biotherapeutics
The impressive advantages of MST, namely the low sample consumption, the broad application range, and swift assay development make it a unique biophysical method…In some cases, LabelFree MST allowed us to perform assays with otherwise “very ill” behaved proteins which were not amenable to any other biophysical technique. Generally, we find very good consistency between quantitative MST measurements and results stemming from other biophysical methods.
Dr. Markus Zeeb
Research Laboratory Head
Boehringer Ingelheim Pharma GmbH & Co.
KG, Germany
DuPont Industrial Biosciences recently acquired its first Prometheus NT.48 nanoDSF instrument. We went through an extensive head to head comparison among other similar technologies and the nanoDSF stood out like a champion. We quickly fell in love and made it the new workhorse in our lab. The instrument provides great data quality of denaturation events with impressive signal to noise ratio, high density of data points, and extraordinary reproducibility making day to day analysis consistent and trustworthy. In addition, the instrument is straight forward to use and data very simple to analyze. We literally, got it, plugged it and started playing in no time.
We started employing the Prometheus to characterize protein samples in early stages of development as well as evaluating formulation stability of super concentrated protein solutions. The instrument is very versatile since measurements require very small sample volume, minimal effort in sample preparation and can handle relevant industrial protein concentrations of up to ~200 mg/mL. In this context, we appreciate the easy sample handling since samples are loaded directly from stock solutions into self-filling capillaries avoiding time consuming and complicated sample preparation steps, even for ultra-concentrated samples. Denaturation events missed by other DSF-type measurements, especially with super concentrated protein sample solutions, are clearly revealed through the high density of data points, yielding same data quality as the gold standard DSC.
Overall, we are very pleased and impressed with the many great qualities of the Prometheus. It will certainly allow us to bridge the gap between molecule screening, selection and product development. We are grateful to NanoTemper for creating this unique biophysical tool as well as for their outstanding and one of a kind customer service.
Dr. Mariliz Johnson
DuPont Industrial Biosciences
Palo Alto, CA USA
I put every protein in Tycho first. The data is very reproducible in our hands. Just put your samples in the capillaries and 3 minutes later you have your results on protein quality. The user interface is genius. You can even enter your experiment labels while it is running to save even more time. It is the only equipment I would recommend to anyone working on proteins because there are no downsides in having a Tycho.
Dr. Stefan Gajewski
Scientist I
Nurix Inc.
MST has proved to be particularly useful to look at molecular interaction involving proteins that are difficult to express or purify. MST requires a relatively short time to setup new assays and is a powerful technique for buffer optimisation. Using the NT.115 MST instrument we have successfully measured small molecule-protein and protein-protein interactions in complex media. Finally, LabelFree MST is one of the few true label-free/immobilisation-free instruments capable of measuring molecular interactions.
Dr. Nicolas Baase
Group Leader, Biochemistry
Sanofi R&D France
This is the first MST instrument we’ve purchased, and we have a lot of confidence in the technique. [In fact], we’re the first people in the CRO world to invest in [NanoTemper’s] capabilities.
Dr. Trevor Perrior
CSO
Domainex
Crelux routinely uses MST as an orthogonal assay for high content hit characterization. Fast assays set-up and straightforward characterization of compound affinity in solution enable us to concentrate on the most promising hits for co-crystallization with their targets and significantly speed up the drug discovery process.
Dr. Ismail Moarefi
Chief Scientific Officer
CRELUX
Martinsried, 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
We have used our Nanotemper NT.115 MicroScale Thermophoresis (MST) instrument extensively in the last one and a half years to study many different types of interaction: protein-small molecule interactions (biological ligands and compounds from medicinal chemistry), small molecule competition assays, protein-metal ion interactions, protein-protein interactions, antibody-antigen interactions and protein oligomerisation. The Facility’s staff and users have benefitted greatly from the excellent customer support provided by Nanotemper. Advice is always practical, relevant and often delivered in-person. Nanotemper’s Applications Scientists are professional, knowledgeable and display an impressive commitment to the success of new users’ experiments.
Dr. Timothy Sharpe
Head of the Biophysics Facility
Biozentrum of the University of Basel
Basel, Switzerland
MicroScale Thermophoresis (MST) is an extremely valuable tool to assess and quantify interactions. In one study using MST, we determined the binding affinities of a wild-type protein and 11 mutants in a half day; this would have been impossible with other techniques. We are using the Monolith NT.115 in our daily research work and we are very satisfied by the data obtained with this novel biophysical method. In addition, we appreciate the support and the competence from the NanoTemper Technologies Company.
Dr. D. J. Hart
European Molecular Biology Laboratory (EMBL)
Grenoble, France
With MicroScale Thermophoresis (MST) we found the ideal alternative and we were immediately convinced by the low sample consumption and immobilization-free setup. Over the last years, we could gain a lot of essential information from our MST experiments enabling us to publish these comprehensive mechanistic insights together with high-resolution crystallographic data. Finally, I would like to emphasize the customer-oriented attitude of this company. Their support team always has an open ear for our questions and offers help with challenging experiments. The company also appreciates our feedback on our experience with the technology and the instruments.
Prof. Dr. Clemens Steegborn
University of Bayreuth
Bayreuth, Germany
Our recent research has focussed on the interactions of proteins involved in the Hedgehog signalling pathway. We have used MST to develop a binding assay with fluorescently labelled peptide and unlabelled protein. The speed of the assay and low sample consumption has allowed us to quickly identify the critical residues in the binding site by rapidly screening a series of mutated proteins. Furthermore, the assay can easily be adapted to screen peptides and small molecules for competitive binding.
Dr. Luca Jovine
Karolinska Institutet
Huddinge, Sweden
In addition to various proteomics technologies and applications, we started to integrate MicroScale Thermophoresis into our portfolio of methods we use to understand DNA repair mechanisms. Using MicroScale Thermophoresis, we could obtain high-quality data on various interactions. We appreciate that the method allows us to optimize our assay conditions quickly in a very straight forward way. The versatility of the method has also proven advantageous in a lab with multiple research projects. We are very pleased by the customer-oriented attitude of NanoTemper Technologies providing valuable user support both in email replies as well as with data discussions in our lab.
Prof. Geir Slupphaug
Norwegian University of Science and Technology (NTNU)
Trondheim, Norway
We are applying MST to determine structural requirements for recognition of complex polysaccharides and the role of ligand-receptor interactions in the relationships between different cells and organisms. MST is very useful for my lab since it allows to measure interactions in solution even in complex samples of membrane proteins. Also the small amount of sample material needed and the broad range of applications are very advantageous.
Prof. Jens Stougaard
Director of CARB
Aarhus University
Denmark
The Core facility at the Weizmann Institute of Science, was created to serve both academic institutions and industrial companies. Therefore, the projects we manage are challenging and diverse. We have three Monoliths to measure biomolecular interactions. All the instruments provide helpful results and are used daily. We also use nanoDSF with the Prometheus NT.48 for the assessment of protein stability before we start any MST experiments.
We would like to point out that we get a lot of support from NanoTemper when more challenging and complex systems are investigated. Overall, MST and nanoDSF are valuable and key techniques in our facility.
Dr. Yael Fridmann-Sirkis
Lab Manager, Life Science Core Facilities
The Weizmann Institute of Science
Rehovot, Israel
MicroScale Thermophoresis (MST) is a quick and easy-to-use biophysical method offering quantitative information on biomolecular interactions in solution. Much of what immediately drew us to the method was immobilization-free approach, fast setup, low sample consumption and maintenance-free instrument – things we appreciate every day using this technology. We have been successfully using Monolith NT.115 to measure small molecule-protein binding affinities from the day one. We find the method a good alternative to our standard enzymatic assays and invaluable approach to proteins without enzymatic activity. We are appreciative of all MST instrument readouts, which allow us not only for Kd determination but also to control the sample aggregation and learn more about protein-ligand binding mechanism. We consider the MST a highly valuable method and recommend it to anyone seeking very sensitive solution-based binding assay.
Dr. Lukasz Krzeminski
Department of Biology
OncoArendi Therapeutics
Warsaw, Poland
Our Monolith NT.115 instrument is a fundamental piece in a set of equipment we use regularly to characterize protein-protein, protein-small molecule and protein-lipid interactions. In order to obtain reliable estimates of binding strength and stoichiometry, we regularly utilize quartz crystal microbalance (QCM), surface plasmon resonance (SPR), and isothermal titration calorimetry (ITC) techniques. MicroScale Thermophoresis (MST) is indispensable for us to measure precious or labor-intensive samples not suitable for ITC because of the required quantity and purity. We found MST suitable for fragment-based design and quality control of biologics development as well. We plan to expand our instrument range with Monolith NT.LabelFree and Seismos in the future. We are very satisfied with the support of NanoTemper and the competence of the company’s application specialists.
Dr. Karoly Liliom
PI
Institute of Enzymology
Research Centre for Natural Sciences
Hungarian Academy of Sciences
Budapest, Hungary
MicroScale Thermophoresis (MST) is our newest addition to this battery of tools for determination of affinity constants, the one for which we hold high expectations with respect to the data quality, speed of determination and low material consumption. Our current MST measurements of binding of Aβ peptide binding to albumin which confirmed the binding constant we obtained previously using CD, took us just a couple of days to complete. MST technology will expand our scientific portfolio in ongoing and future projects.
Prof. Dr. hab. Wojciech Bal
Polish Academy of Sciences
Warsaw, Poland
In my lab, MicroScale Thermophoresis NT.115 by Nanotemper opened an important new avenue how to get rapid data on protein/protein interactions in difficult receptor-ligand systems independently of the methods measuring the interactions on surface such as Surface Plasmon Resonance.
Dr. Bohdan Schneider
Academy of Sciences of the Czech Republic
Prague, Czech Republic
Since we are specialized in the study of biomolecular interactions, recently acquired Monolith NT.115 nicely complements other instruments operated in our Core Facility (e.g. calorimeters, SPR biosensors, analytical ultracentrifuge). Due to the low sample consumption and short time required for the setup of the experiment, we have been using MicroScale Thermophoresis mainly as a preliminary screening method for potential ligands of the biomolecules tested. We have encountered rare cases of proteins where the aforementioned techniques failed due to protein precipitation and MST was the only applicable technique to get Kd of the interaction. I find the MicroScale Thermophoresis a valuable tool for the study of interactions and I also appreciate the excellent customer services done by the representatives of NanoTemper Technologies. They are always very helpful in terms of experimental setup and data analysis.
Dr. Michaela Wimmerová
CEITEC
Brno, Czech Republic
In my group we explore the structural, biophysical, and cellular outcomes of protein complex formation at membrane-bound receptors. We use Microscale Thermophoresis (MST) in addition to other methods including isothermal titration calorimetry and surface plasmon resonance and find a good agreement between the methods. We are very pleased with the low material consumption, short measurement times and broad application range of MST.
Prof. John E. Ladbury
Astbury Centre for Structural Molecular Biology
University of Leeds
United Kingdom
We are very enthusiastic about MST as it quickly enabled us to measure protein:protein and protein:small molecule interactions that have been difficult to detect and quantify with standard biomolecule interaction technologies in the past years. MST provides us with a unique tool to validate the lead inhibitors that bind to specific sites of Rho GTPases or their regulators/effectors.
Dr. Yi Zheng
University of Cincinnati College of Medicine
OH, USA
At ChromoTek, we develop, manufacture, and market innovative research tools based on alpaca antibodies; these reagents and kits improve research applications in both biochemistry and cell biology. NanoTemper Technologies’ Prometheus NT.48 instrument has quickly become an integral part of ChromoTek’s R&D workflows as it allows us to quickly obtain critical data on protein characteristics and stability early within the development process. We also use the Prometheus routinely for the QC of our high-quality products like the GFP-Trap®. Our scientists are excited by the Prometheus’ ease of use, low sample consumption, and reliability.”
Dr. Felix Hartlepp
VP Research & Technology
ChromoTek GmbH
Martinsried, Germany
The Monolith NT.115 instrument works great both in terms of ease of use and short time for setting up the experiment. The sample consumption is very low and costs are cheap. Its immobilization-free approach makes things less complicated in terms of handling protein samples and setting-up the assay. The new version of the software for analyzing the results shows large improvement over the previous version. The technical support and customer service from NanoTemper are excellent from the very first days of the installation of the instrument. Overall, we are very satisfied with it and we strongly recommend it to research groups looking for fast quantitative binding assay.
Prof. Antonio Macchiarulo
Department of Pharmaceutical Sciences
University of Perugia
Italy
We study the structural and molecular biology of bacterial adhesins and cell-surface filaments with respect to their function in bacterial pathogenesis, with the ultimate aim of developing a new generation of virulence-targeted antimicrobials. MST really is opening up the easy determination of some Kd’s that were hard to get to with other methods. So far, we measured protein-protein, protein-glycan, protein-small compound and protein-cofactor interactions with the Monolith NT.115.
Prof. Han Remaut
Vrije Universiteit
Brussel, Belgium
Microscale Thermophoresis (MST) is an extremely valuable tool to assess and quantify ligand-receptor, cofactor-receptor and nucleic acid-receptor interactions. […] In addition to ITC, MST allowed us to examine a range of concentration ratios between the two binding partners that could not be reached with ITC and to demonstrate the existence of allosteric control in the binding of the coregulator to the homodimeric nuclear receptor. We are using the Monolith NT.115 in our daily research work and we are very satisfied by the data obtained with this novel biophysical method. In addition, we appreciate the support and the competence from the NanoTemper Technologies Company.
Dr. Isabelle M.L. Billas
Institute of Genetics and Molecular and Cellular Biology (IGBMC)
France
Our main focus is looking for small molecular inhibitors for receptors like PD-L1. We have found MST to be useful to determine drug potency due to its versatility and low sample size requirement. The results correlate nicely with the ones obtained from other techniques (HTRF and Fluorescence Polarization Assays), but thanks to an instrument with low complexity like Monolith, we find MST to be more reliable. Lastly, I’d like to highlight the software as very user friendly, with an interface that guides you through the whole procedure and readily analyses the results.
Dr. Jacek Plewka
Post-doc
Jagiellonian University, Department of Chemistry
Kraków, Poland
With the new MST technology we were able to measure the interaction of membrane vesicles, mediated by coiled coil-forming peptides. The technology requires only little sample material and is enabling for this type of experiments.
Prof. Dr. Andreas Janshoff
Georg-August-Universität Göttingen
Germany
We are interested in the biophysical properties of soluble and membrane bound biomolecules and are using MST to measure binding affinities and complex formation. MST is straightforward and fits well in our technology portfolio to cover a broad spectrum of synthetic biomolecules including proteins, peptides and nucleic acids.
Prof. Dr. Ulf Diederichsen
Georg-August-Universität Göttingen
Germany
To investigate biomolecular interactions, my group uses a wide range of techniques. MicroScale Thermophoresis (MST) is mainly used as a fast approach for the initial characterization of interactions. Advantages include low material consumption, short measurement times and a wide range of accessible interactions. Kd values obtained with MST are generally in good agreement with those obtained using ITC or Ki values determined with the help of enzymatic assays.
Prof. Dr. Dietmar Manstein
Hannover Medical School
Germany
Our lab focuses on the importance for telomere and telomerase to maintain the integrity of genomes and the stability of chromosomes. Large amount of proteins and protein complexes are involved in epigenetics, which play essential rotes in controlling many cellular events. We are employing MicroScale Thermophoresis (MST) in determining the interactions of proteins/protein complexes to ssDNA/RNA. It’s easier to measure the protein-protein or protein-nucleic acid interaction, especially for those sensitive samples, that has been difficult to detect and quantify with ITC or SPR. We benefited from the low sarnple consumption, the fast measurements, the broad application range and the possibility to choose the buffer freely, resulting in highly reproducible data. We strongly recommend MST technology for its low material consumption, short measurement times and very high sensitivity.
Prof. Dr. Lei Ming
Director, National Center for Protein Science Shanghai (NCPSS)
Deputy Director, Shanghai Institute of Biochemistry and Cell Biology (SIBCB)
Shanghai, China
For such weak interactions [millimolar to micromolar], it is difficult to set up reliable biochemical assays or binding assays using ITC or SPR techniques. Fortunately, MST technology turns out to be an excellent way to determine such weak interactions between small fragments and protein in our system of interest. We efficiently and reliably performed our experimental binding measurement using minimum amount of protein and fragments. We strongly recommend MST technology for its low material consumption, short measurement time, and very high sensitivity.
Dr. Niu Huang
Associate Investigator
NIBS
Beijing, China
We used Microscale Thermophoresis (MST) to measure the interaction between large molecular chaperones with molecular weight up to 800 kDa. It is distinguished in low sample consumption, fast measurement and immobilization-free setup. We found that the biophysical data obtained from MST are in good agreement with those obtained from SPR or ITC. MST technology will facilitate our research on various interactions.
Prof. Cuimin Liu
Institute of Genetics and Developmental Biology
Chinese Academy of Sciences
China
For the Biochemistry Core Facility at the Max-Planck-Institute for Biochemistry it is of great importance to provide our users with instruments which are easy to use while offering a high information content. Therefore, the NanoTemper Monolith NT.115 with its low sample consumption and maintenance-free design is a perfect addition to our instrument park. The straight forward handling of the instrument allows even first time users to access binding affinities usually within a day. Method development is often quick, certainly compared to other techniques, but the method puts a high demand on sample homogeneity and in particular on monodispersity. As interactions are characterized free in solution, MST has been successful where SPR failed due to the limitations of a surface based approach.
Dr. Stephan Uebel
Max-Planck-Institute of Biochemistry
Martinsried, Germany
MicroScale Thermophoresis (MST) by NanoTemper Technologies is very useful tool to quantify biomolecular interactions. The advantage of this platform is that the assay is simple to set up, it requires minimal reagents and biomolecular interactions can be measured under native conditions with minimal alterations to reactants. At Center for Chemical biology and Therapeutics, inStem, National center for biological sciences (NCBS), we have been using MST routinely to measure the protein-small molecule interactions across multiple protein domains successfully as an alternative to rather cumbersome approaches such as ITC, SPR and TSA. It is quick, precise and highly informative in reading the molecular interactions.
Dr. Muralidhara Padigaru
Institute for Stem Cell Biology and Regenerative Medicine
Bangalore, India
Via MST, we measured the Protein-DNA interaction, that have been difficult to detect and quantify with other standard biomolecule interaction technologies (like SPR and ITC) because of the conformation/immobilization-sensitive and very little heat changes of our samples. MST technology can be used as a complementary technology to the traditional methods in these fields. We can strongly recommend MST technology for its low material consumption, short measurement times and broad application range for molecule-interaction studies.
Prof. Dacheng Wang
Chinese Academy of Science
China
It has proved a valuable tool for characterising small molecule-protein and protein-protein interactions, as well as for the study of protein aggregation concentration determination. There is very good agreement with other technologies such as Surface Plasmon Resonance (SPR) and Isothermal Titration Calorimetry (ITC), and we are particularly appreciative of this new technology because of the extremely low protein consumption and relatively short time required for the assay setup.
Dr. Alexey Rak
Head, Bio Structure and Biophysics
Sanofi R&D France
We measured the dissociation constant for periplasmic binding protein and siderophore through MST technology. Because siderophore is extremely insoluble, it’s difficult for us to complete that task via other methods such as ITC and SPR. We strongly recommend MST technology for its low material consumption, short measurement time and very wide application range.
Prof. Li-chuan GU
Shandong University
China
Our company 2bind works with worldwide customers from pharma and biotech companies as well as with universities on the characterization of molecular interactions, mainly in the areas of drug development, antibody discovery and aptamer design. The MicroScale Thermophoresis technology allows us to offer cost-efficient, precise and fast analyses of biomolecular interactions of any kind. The integrated quality controls of the MST system enable us to easily identify sticking and aggregation effects and to directly improve measurement conditions ensuring high quality data. Our services based on the MST technology are thus perfectly suitable for the identification and characterization of molecular interactions.
Dr. Thomas Schubert
CEO
2bind GmbH
Germany
We use MicroScale Thermophoresis (MST) to measure the affinity of our compounds, aptamers, and nanodiscs to their targets. Using this technology, we obtained very convincing data that were in good agreement with results obtained from ITC and NMR. Its advantages include low material consumption, short measurement times, immobilization-free setup, maintenance-free instrumentation, and a wide range of accessible interactions. MST is mainly used as an easy and fast approach for the characterization of interactions.
Prof. Liu Qingsong
High Magnetic Field Laboratory
Chinese Academy of Sciences
China
In order for proteins to manifest their proper biological and therapeutic effect, their conformational and structural integrity must be maintained at all stages of the development and commercialization process. Thus, for the development of Biotherapeutics it is essential to find optimal (solution) conditions to stabilize the Drug Product such that we can ensure its safety, biological function, structural integrity, and storage for long term under unfavourable and favourable conditions.
To meet the requirements of current clinical indications with respect to drug delivery, highly concentrated therapeutic protein drugs up to 100-200 mg/ml are necessary. In order to achieve such concentrations, a number of protein properties like solubility, self-association, solution viscosity, and protein aggregation have to be controlled. The large dynamic range of the Prometheus NT.48 allows for analyzing thermal unfolding and re-folding in solutions containing protein concentrations between 250 mg/ml down to few µg/ml. Moreover, measurements with rather viscous sample solutions as sometimes observed by highly concentrated formulations can be performed. Thus, it can be utilized for both, stability screening during early phases of drug discovery where only small amounts of protein are available, as well as for formulation screening campaigns of highly concentrated samples. The capabilities of the Prometheus NT.48 itself and in combination with other complementary biophysical tools enables for a better understanding of conformational and colloidal properties of the protein candidate and their impact on macroscopic solution properties. However, in some cases, the Prometheus NT.48 allowed us to perform thermal stability screening with otherwise structurally very complex proteins which were virtually not amenable to any orthogonal biophysical technique.
Moreover, the Prometheus instrument allows for label-free analysis of 48 samples simultaneously independent of their protein concentrations (high dynamic range) and selected solution conditions and/or –compositions. Unlike other techniques the Prometheus NT.48 measurements remained unaffected by any excipient, sugar, detergent or additive. Altogether, the Prometheus instrument enables for very flexible experimental design and provides maintenance-free instrumentation. In addition, our obtained data demonstrate very high reproducibility, consistency, the robustness and precision of this particular technology. The outstanding construction design allows for on-the-fly detection of fluorescence intensity resulting in impressive data point density that there is virtually no need for data fitting.
I personally also highly appreciate the very intuitive software allowing for rapidly setting up the experiment and evaluation of obtained data. The software being developed allows for rapid/easy data analysis, data interpretation and exportation to other data processing software.
Dr. Michaela Blech
Associate Director
Boehringer Ingelheim Pharma GmbH & Co. KG
Biberach, Germany
See why OncoArendi Therapeutics chose Monolith MST for measuring binding affinities
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 Kd. 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 Kds 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.
Dr. Xavier Thuru, Scientific Manager
Romain Magnez, Technical Manager
Dr. Isabelle Van Seuningen, Laboratory Director
Canther
Lille, France