新的分子间相互作用分析仪微量热泳动仪---MST 用户评价举例

Our recent research is focused on the molecular basis of herbicide resistance and exploiting new target sites of herbicide, such as PPI of AHAS. Via MST, we measured the Protein-Protein interaction, that have been difficult to detect and quantify with ITC because of instability of the protein and very little heat changes of our samples. 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.

我们目前致力于除草剂抗性的分子机制以及寻找新的除草剂靶点（比如AHAS亚基间相互作用的界面）等方面的研究。通过MST，我们测量了蛋白质-蛋白质相互作用，而我们的样品使用ITC方法测定没有得到结果，因为我们的样品稳定性较差并且相互作用热量变化极小。所以在这些研究领域，MST技术能够很好地与传统的技术互补。

王大成

Our resent research is focused on the critical pathogenic protein & complex from pathogenic microbes and The Protein structure-functional basis for some endogenous disease. 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.

我们非常推荐MST技术的一些特点，如低样品使用量、很短的测量时间以及在分子间相互作用方面很广泛的应用。

谷立川

Part of our research focus on the protein structural-function basis for iron metabolism of pathogenic microorganisms. 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.

我们目前一部分课题是关于致病微生物铁代谢相关蛋白的结构与功能。通过MST技术，我们测量了周质空间转铁蛋白与铁载体的解离常数，我们的样品很难通过其他传统方法（例如SPR和ITC）来测量因为小分子的溶解度极低，MST技术的我们得到了理想的结果。我们非常推荐MST技术的一些特点，如极低的样品使用量，较快的测量时间，以及十分广泛的应用。

“We routinely assess interaction affinity for both small molecule and biologics projects, with NanoTemper Technologies’ Microscale Thermophoresis being the most recent addition to the pool of instruments we use to carry out these measurements. 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. NanoTemper customer support has been a key factor in enabling us to familiarise ourselves with the new technology. We would like to deploy increasing numbers of applications based on MST technologies and continue to interact with NanoTemper Technologies Company, a dynamic, scientifically driven company.”

（编者译）

我们经常需要测量小分子和生物制剂的相互作用亲和力，我们最近使用的最新一个设备就是NanoTemper的微量热泳动技术。结果证明了MST是一个有价值的工具，用于测量小分子-蛋白质、蛋白质之间的相互作用，以及判断蛋白质形成聚集体的浓度。MST技术与传统的其他技术如表面等离子体共振(SPR)和等温滴定量热法(ITC)的结果完全吻合，我们特别喜欢这个新技术的一些特点：极低的蛋白样品用量和很短的实验设置时间。NanoTemper的技术支持非常及时有效，使我们能够很快熟悉这个新技术。我们很愿意使用越来越多MST仪器并继续与NanoTemper技术公司相互合作，因为这个一个积极向上的、技术先进的公司。

 

（编者译）

我们的主要工作是根据结构设计药物。比如，细菌tRNA修饰酶-tRNA鸟嘌呤转糖苷酶(Tgt)与小分子抑制剂以及tRNA的相互作用。虽然我们无法直接衡量这个酶的与小分子鸟嘌呤以及7-deaza-7-aminomethyl的相互作用，但是由于Tgt结合大分子tRNA是有很明显的热泳动效应变化，我们可以通过MST来测量各自的Kd值。此外，MST测量出来lin-benzoguanine-based MST Tgt抑制剂(300 - 500 Da) 的Kd值极好地吻合先前确定通过酶动力学研究得到的Ki值。因此，这个快速和易于使用的方法提供了一个非常好的方法用于替代放射性酶活性测定，而后者是我们之前一直用来测量Tgt抑制剂亲和力的方法。我们使用到目前为止找出绑定的亲和力Tgt抑制剂。此外,我们还研究了一个志贺氏杆菌III型分泌系统分子伴侣蛋白与其结合物（合成的多肽）的相互作用。通过MST，我们能通过极低量的蛋白质来测定Kd值，而且其结果几乎符合传统的等温滴定量热法ITC等技术所得到的数据。

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 rapidly quantify relevant protein-nucleic acid and protein-protein interactions directly in cellular lysates without going through the hassles of purification.

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.

 

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.”

 

We are studying the molecular mechanisms of signal transduction processes involving Rho GTPases and are developing small molecule inhibitors and other strategies that interfere with specific Rho protein functions in leukemia, lymphoma and lung cancer.

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 .

（编者译）

我们正在研究Rho GTPases的信号转导的分子机制的，并开发小分子抑制剂和其他方法用于特异性地干扰白血病、淋巴瘤、肺癌中的Rho蛋白质功能。

我们很喜欢MST技术，因为在过去的几年里MST帮助我们快速完成测量蛋白质-蛋白质和蛋白质-小分子间的相互作用，很多时候很难使用其他标准的生物分子互作技术完成。MST为我们提供了一个独特的工具来验证重要的抑制剂是否结合到Rho GTPases的特异性位点或者结合到他的调节物/效应物。

 

“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”

“We are interested in understanding the interactions between cells and organisms by investigating the role of polysaccharides exposed on cell surfaces and secreted polysaccharide signal molecules. 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.”

（编者译）
我们的实验室通过研究暴露在细胞表面的多糖和分泌型多糖信号分子来揭示细胞与细胞以及有机体的相互作用。我们使用MST来测量复杂的多糖的结合以及配体-受体的结合以阐明不同细胞或机体间的相互作用。

My lab focuses on the elucidation of the structure and function of membrane proteins. This topic includes the investigation of protein-protein and protein-substrate 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.

（编者译）

我们的实验室专注于研究膜蛋白的结构和功能，包括蛋白-蛋白以及蛋白-底物相互作用的研究。我们使用MST来研究膜蛋白感应器分子-传感器分子-转录因子-DNA的结合反应。MST显示出极好的优点，包括快速和简单测量，样品消耗量很少等等。

Our lab focuses on the mechanisms and organization of DNA packaging inside the cell nucleus. We study the biochemical properties of molecular machines, the chromatin remodelers that regulate DNA accessibility and switch between ‘on’ and ‘off’ states of genes.

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.

（编者译）

我们的实验室专注于研究细胞和之中的DNA包装的机制，研究分子机器的生化特性，染色质重塑调节DNA组装和调节基因的开关状态。我们通过MST来研究染色质重组化酶并且定量他们与DNA，RNA以及其他蛋白之间的亲和力。MST将定量生化技术推向一个新的台阶因为其简单易学，并且非常好的取代了传统的EMSA技术。

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.

（编者译）

我们对实验胚胎学的定义和维护组蛋白准则的基本机制很感兴趣，并且使用MST来测量“Reader”蛋白和修饰后的组蛋白多肽的结合以及“writer”酶（包括激酶、去甲基化酶、甲基化酶）的激活剂和抑制剂作用。

The work of our group gave rise to the novel concept that under certain conditions matrix components may act as endogenous “danger” signals, which are recognized by innate immunity receptors, and are capable to trigger an inflammatory response reaction.

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.”

（编者译）

我们发现在特定化井下某些基质成分会变成内源性的“危险”信号，而这些异常的成分会被先天免疫受体识别，并且能够引发炎症反应。

我们使用MST来测量细胞外基质中的内源物/体内突变的蛋白聚糖与他们相关受体的反应，并且非常满意目前获得的所有数据，所得到的数据与其他生物方法预测的完全吻合并且验证了之前使用免疫沉淀和结合试验所获得的数据。

MST是一种非常新的技术，我们推荐这个技术可以用来获得非常好的定量的亲和力数据。

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.

（编者译）

我们正在研究果蝇前体细胞的MicroRNA发生过程，并且使用MST来测量核酶即特异性的因子结合到RNA上的反应。MST极少的蛋白样品的消耗量和极短的测量时间使得他成为一个非常有价值的技术，它能帮助我们回答很多之前无法解释的问题。

Membrane interaction and fusion plays a fundamental role in many cellular processes such as intracellular trafficking, fertilization, tissue formation or viral infection.

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”（编者译）

生物膜的融合以相互作用在细胞的多项过程中（细胞内运输，受精，组织形成以及病毒感染）起到非常重要的基础作用。通过MST技术，我们能够测量螺旋状多肽介导的囊泡的相互作用。该技术仅仅需要极少的样品材料即可完成测量。

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.

（编者译）

生物膜的融合以相互作用在细胞的多项过程中（细胞内运输，受精，组织形成以及病毒感染）起到非常重要的基础作用。通过MST技术，我们能够测量螺旋状多肽介导的囊泡的相互作用。该技术仅仅需要极少的样品材料即可完成测量。我们对游离的和膜上生物分子的生物物理学性质感兴趣，并且使用MST来测量结合亲和力和复合物的构象。

 

 

We are interested in molecular architectures especially to stabilize and analyze challenging smell receptors that belong to G protein-coupled receptors (GPCRs). GPCR is a super-family of receptors linked to many diseases. We want to measure the interactions of smell receptors with odorant that are very low molecular weight ligands. In the past that has been extremely difficult to detect and quantify other common molecular interaction technologies including SPR, and nearly impossible to use ITC for odorant-binding measurement because of the conformation and immobilization-sensitivities, or difficult and costly to obtain sufficient amount of receptors. 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.