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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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

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.

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.

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.

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.

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.” 

The nanoDSF instrument from NanoTemper Technologies has become one of the primary techniques to screen for the thermal stability of globular proteins at Novo Nordisk A/S. The instrument and software are intuitive and easy to handle which makes this suitable as a walk-up instrument. The method is very versatile due to fluorescence and aggregation optics, low sample consumption, concentration range and insensitivity towards many buffers and excipients. For globular proteins with tryptophan residues we yield high quality thermal unfolding data which compare with other conventional methods.

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.

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.

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.

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.

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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.

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).

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

or one and a half years we have offered MicroScale Thermophoresis and it has become the most frequently used technique to study biomolecular interactions for scientists from the Weizmann and from other academic institutions and companies in Israel. Particularly important is the fact that MST can be used to study many different types of interaction. A very valuable advantage of MST is a significantly lower percentage of false positive (unspecific) results compared to other biophysical techniques. In addition we appreciate the straight forward handling and the low sample consumption.

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.

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. 

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.

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.

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.

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. 

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.