Customer Statements
MicroScale Thermophoresis
“
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, Boehringer Ingelheim Pharma GmbH & Co. KG, Germany
“
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, Sanofi R&D, France.
“
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, Actelion Pharmaceuticals Ltd. Allschwil, Switzerland
“
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 moleculeprotein 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 Basse, UCB-Celltech, UK
2
MicroScale Thermophoresis
“
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 maintenancefree 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
“
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, University of 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
“
Often, fluorescent microscopy, employing tagged or labeled microtubule
associated proteins (MAPs) and motors, is used to study tubulin assembly and
microtubule based motility.
Previously, we have employed size exclusion chromatography, sucrose gradient
sedimentation and immuno-precipitation to detect but not quantify these
interactions as the necessary instrumentation was unavailable. MicroScale
Thermophoresis has proven an ideal way for cell biologists in our institute to
quantify interactions as, in most cases, the fluorescently tagged protein
preparation is already available and, most importantly, only a limited amount of
sample is required.”
Dr. David Drechsler, Max-Planck-Institute of Molecular Cell Biology and Genetics, Dresden, Germany
3
MicroScale Thermophoresis
“
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, Germany
“
In vitro interaction analyses are a simple but very informative tool, which
can readily use the recombinant protein already available. Compared to more
traditional methods such as Isothermal Calorimetry (ITC) and Surface Plasmon
Resonance (SPR), MicroScale Thermophoresis (MST) stands out due to its ease
of use, its fast setup and its very low consumption of biomolecules. We consider it
a highly valuable alternative.
Prof. Dr. Thomas Müller, Julius-von-Sachs-Institute, University of Wuerzburg, Germany
“
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, Germany
“
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 of 2bind GmbH, Germany
4
MicroScale Thermophoresis
“
My group at the Universitätsklinikum Hamburg-Eppendorf is interested in the
development of nanobodies. Nanobodies can be easily produced as recombinant
proteins and are therefore interesting molecules for pharmaceutical research. We
are using MicroScale Thermophoresis (MST) to characterize the binding affinities of
nanobodies to their antigens. We appreciate that MST is capable of measuring
antibody-antigen interactions in solution. Nanobodies bind to their antigen with
affinities in the low nanomolar range, which can be precisely quantified by MST.
Other advantages of this approach include simple handling and maintenance, low
material consumption, and fast analysis.”
Prof. Friedrich Koch-Nolte, Universitätsklinikum Hamburg-Eppendorf, Hamburg, Germany
“
My lab focuses on the elucidation of the structure and function of membrane
proteins. This topic includes the investigation of protein-protein and proteinsubstrate interactions. We are employing MST in binding studies of membrane
sensors to their transducers as well as soluble transcription factors to DNA. We also
plan for investigations of receptor – ligand interactions with this exciting new
technique. MST turned out to be an extremely useful method in our lab to determine
binding constants. It is advantageous that the fast and handy experiments require
only few amounts of protein.”
Prof. Dr. Joachim Heberle, FU Berlin, Germany
“
MST yielded Kd values for lin-benzoguanine-based Tgt inhibitors (300500 Da), which were excellently consistent with Ki values previously determined in
enzyme kinetic studies. Accordingly, this fast and easy to use method provides a
highly welcome alternative to the radioactive enzyme assay we used so far to
figure out binding affinities of Tgt inhibitors. […] Via MST, we determined with low
amounts of protein material Kd values which were in nearly perfect agreement with
those measured via isothermal titration calorimetry.”
Prof. Dr. Gerhard Klebe, Philipps-Universität Marburg, Germany
“
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, Germany
5
MicroScale Thermophoresis
“
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
“
The MicroScale Thermophoresis (MST) technology is a good complement
to other biophysical techniques we are applying in the lab to investigate receptor
proteins involved in inflammatory response. However, MST provides additional
benefits since we are interested in elucidating interactions in close-to-native
conditions: thus, we appreciate that the interactions are investigated free in
solution and can be studied even in cell lysate. We are using MST for a number
of different interactions, mainly to investigate protein-protein interactions but also
to study protein-DNA interactions and protein binding to ions.
MST is a novel l technique which is not related to any other established biophysical
methods thus providing independent insights into biomolecular interactions. It is
very easy to use with low time and material consumption.”
Prof. Dr. med. Uffe Holmskov, University of Southern Denmark, Odense, Denmark
“
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
6
MicroScale Thermophoresis
“
For 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.”
Dr. Aharon Rabinkov and Dr. Irina Shin, Weizmann Institute of Science, Rehovot, Israel
“
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 fragmentbased 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, RCNS, HAS, 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
7
MicroScale Thermophoresis
“
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
“
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.“
Prof. Dr. Michaela Wimmerová, CEITEC, Brno, Czech Republic
8
MicroScale Thermophoresis
“
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, The University of Texas, Houston, TX, USA
“
The MST technology is a superb technology, it not only is label-free, hence
reducing the measuring artifacts, but it also is surface-free that provides true
measurement of finest molecular interactions. Thus it allows us much better
analysis of the GPCR with their small-molecule ligands. We strongly recommend
MST technology for its low material consumption (a few micrograms in a few
microliters), short measurement times, inexpensive consumables, and broad
application range in all molecule-interaction study.”
Dr. Shuguang Zhang, Massachusetts Institute of Technology, Boston, MA, USA
“
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
“
“
To date, my laboratory has enjoyed considerable success in using
NanoTemper instruments to precisely determine the affinity of biomolecular
interactions (e.g. protein-small molecule, protein-protein and protein-peptide
interactions). Key advantages are rapid optimisation of binding assays, low
material consumption and being able to easily validate putative interactions in vitro
before committing to extensive protein engineering or structural analyses.
Dr. Neil Ferguson, Conway Institute, University College Dublin, Ireland
9
MicroScale Thermophoresis
“
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 use systems biology to study transcription regulatory phenomena on a
genome-wide scale in the context of host-pathogen interactions and cancer.
Microscale Thermophoresis (MST) thereby has been extremely useful to
rapidlyquantify relevant protein-nucleic acid and protein-protein interactions directly
in cellular lysates without going through the hassles of purification.”
Dr. Arndt Benecke, Centre National de la Recherche Scientifique, Paris, France
“
We study the structural and molecular biology of bacterial adhesins and cellsurface 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, proteinsmall 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.
10
MicroScale Thermophoresis
“
In our MST studies, we used the technology to precisely quantify the
cooperativity of PIP2 and calcium binding. [...] Given (i) the relatively high affinity
of synaptotagmin-1 for both PIP2 and calcium, (ii) the low solubility of PIP2 and
(iii) that synaptotagmin-1 is prone to aggregate at high protein concentrations, we
cannot think of any technique other than MST that would allow studying this
cooperative binding to such detail. Moreover, due to the very high sensitivity of
MST, we estimate that the same amount of synaptotagmin-1 suffices to record
about 10,000 times more calcium binding curves with MST compared to NMR or
ITC.”
Dr. Geert van den Bogaart, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
“
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, University of 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
11
MicroScale Thermophoresis
“
We are studying the processing of microRNA precursors In Drosophila and
are using MST to measure the binding of nucleolytic enzymes and their specificity
factors to RNA substrates.
The small amount of protein sample needed and the much faster measuring time
compared with e.g. gel-shift assays are particular strengths of this new technique.
It enables us to ask questions that we could not address before.”
Prof. Dr. Klaus Förstemann, Gene Center of the LMU, Munich, Germany
“
We are interested in the basic mechanisms in epigenetics which define and
maintain the histone code and are using MST to measure the binding of “reader”proteins to modified histone peptides as well as the activity and inhibition of “writer”
enzymes including kinases, demethylases and methylases.
The new MST technology is easy to use and a good alternative to our standard
enzymatic assays which are radioactive and measure endpoints only. It allows us
to easily measure affinities for protein-peptide interactions in solution”
Prof. Dr. Axel Imhof, Adolf Butenand Institute, LMU, Munich, Germany
“
Our lab is developing diagnostic tools based on protein biochips and is studying
the basic mechanisms underlying herpes virus infections” We are using MST for
elucidating the function of viral proteins and its interaction with other viral and host cell
proteins.
MST is simple to use and was quickly established in our lab. In particular we do
appreciate the low instrument, consumables and maintenance costs associated with
this new technology”
Dr. Maria G. Vizoso Pinto, Max-von-Pettenkofer Institute, Munich, Germany
“
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 of CRELUX, Martinsried, Germany.
12
MicroScale Thermophoresis
“
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
“
We measured the binding of several growth factors to CD44 using the
NanoTemper Technologies’ Microscale Thermophoresis. Using this technology we
obtained very convincing data that were in agreement with results obtained using
other technologies like Surface Plasmon Resonance (SPR). This method is
particularly useful to perform low affinity binding measurements using low amounts
of proteins. We are extremely satisfied with the quantitative data obtained with this
new technology.”
PD Dr. Orian-Rousseau, Karlsruher Institut für Technologie (KIT) Germany
“
We are using MST to measure the interactions of endogenous and in vitro
mutagenized proteoglycans of the extracellular matrix with their receptors and
are very satisfied with the results obtained so far. The affinities are consistent with
the biological readouts and confirm previous results gained with
immunprecipitation and binding assays. MST is a new technology we can
definitely recommend for obtaining robust quantitative affinity data.”
Prof. Dr. med. Liliana Schaefer, Pharmazentrum/ZAFES, Frankfurt a.M., Germany
13
MicroScale Thermophoresis
“
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
“
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 proteinnucleic 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 at the National Center for Protein Science Shanghai (NCPSS) and Deputy
Director at the 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 immobilizationfree 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
14
MicroScale Thermophoresis
“
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
“
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
“
We measured the dissociated 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
15
nanoDSF
“
Native, intrinsic fluorescence based nanoDSF technology has been
quickly integrated in our standard operations and is applied nowadays in every
project we are currently working on, including small molecules and biologics
modalities. We are using Prometheus for initial screening as well as for lead
optimization profiling of small molecules and fragments and for stability
characterization of therapeutic proteins. Furthermore, nanoDSF is an invaluable
tool for quality control since the fraction of unfolded protein can be quantified just
within a few seconds.
The Prometheus NT.48 instrument is maintenance-free and provides an easy to
use handling platform. The capillary format allows us to even measure highly
viscous formulation conditions.
We experienced that nanoDSF technology is superior to standard DSF regarding
its application range as well as in terms of precision, reproducibility, wider applicability and greater
potential for new applications development.”
Dr. Alexey Rak, Sanofi R&D, France
“
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. All together, 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
“
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 of Targenomix GmbH, Potsdam, Germany
16
nanoDSF
“
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
“
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
“
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 maintenancefree.”
Dr. Thomas Schubert, CEO of 2bind GmbH, Germany
17
nanoDSF
“
Very few methods exist for membrane proteins, as the requirement of
detergent-containing buffers very often impedes the use of CD spectroscopy
(due to strong light absorption by the detergent) and the application of the
usually preferred ThermoFluor methodology (due to binding of the dye to the
detergent micelles).
The capabilities of the new Prometheus NT.48 in measuring thermal unfolding
now allows us to quickly determine the ideal buffer conditions and the
detergent best suited for crystallization trials. Fast measurements, very low
material consumption, label-free capabilities, and low background noise make
the Prometheus NT.48 the best current solution for screening membrane
protein buffer conditions.”
Prof. Dr. Thomas Müller, Julius-von-Sachs-Institute, University of Wuerzburg, Germany
“
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
“
Over the years, my co-workers and I have measured the stability of many
tens of thousands of proteins (or mutants thereof). Typically, these take the form
of chemical denaturant titrations or thermal denaturation assays. Whilst the latter
has proved possible to automate via different strategies, chemical denaturant
titrations have historically been slow, required user skill and continual
intervention to achieve good results and consumed lots of protein. This has led
to denaturant titrations being a niche, primarily academic, technique. However,
the NanoTemper Prometheus changes this, as it allows very rapid, low protein
consumption determination of protein stability using chemical and/or thermal
denaturation. Thus, the Prometheus is an ideal workhorse for academic research
involving large-scale protein engineering or stability measurements.”
Dr. Neil Ferguson, Conway Institute, University College Dublin, Ireland
18
Surface Acoustic Wave
“
My group is focusing on the characterization of molecular mechanisms
mainly by x-ray crystallography. Amongst other biophysical technologies, we
also employ the Surface Acoustic Wave (SAW) technology for various different
projects. The SAW technology allowed us to decipher the complex interplay of
orthosteric and allosteric inhibition in legumain, which was critical for a
mechanistic understanding of the dual protease and ligase activities of
legumain. We very much appreciate the great service as well as the helpful
and customer-oriented attitude of NanoTemper Application Scientists.”
Prof. Johann Brandstetter, Group Leader Structural Biology Group, University of Salzburg, Austria
“
We realized the higher sensitivity of the SAW technology compared to our
QCM approaches, initially analyzing protein-protein interactions. We established
a technology where well defined model membranes were externally transferred
to the sensor chips, subsequently dried and rehydrated in the sensor device
before the measurement. Using this technology we obtained e.g. an insight into
the complex interplay of cell surface proteoglycans with signaling molecules and
the potential interference by heparin-based drugs in tumor cell metastasis.
The SAW team members were always helpful and if necessary innovative in
supporting us on our particular and technologically challenging tasks.”
Prof. Dr. Gerd Bendas, Pharmaceutical Institute, University of Bonn, Germany
“
Complex systems are to be analyzed, among which my area of
expertise lies on vesicles, liposomes and membrane monolayers and micellar
structures.
The SAW technology was the first biosensor suited to comply with these
versatile applications. From interactions like kinases and nucleotidases with
nucleotides, both the kinetics of interactions can be calculated, and induced
conformational changes can be differentiated. In addition, the SAW biosensor
may open the unique possibility to detect macromolecular complexes
consisting of proteins and (phospho)lipids within complex matrices and thereby
enable the identification of novel disease biomarkers.
We appreciate the competence of the SAW team providing valuable user
support.”
Dr. Gerd Müller, Helmholtz Center Munich, Germany
19
Contact
NanoTemper Technologies
GmbH
Flößergasse 4
81369 München
Germany
Phone: +49 (0)89 4522895 0
Fax: +49 (0)89 4522895 60
[email protected]
http://www.nanotemper-technologies.com
V46_2016-05-20