CryoProbes

BioSolids CryoProbe

Increasing Sensitivity Without Sample Modification

NMR虽然非常通用,但它具有不敏感的固有挑战。提高不需要样品修饰的灵敏度的一种策略是冷冻冷却RF线圈和前置放大器电子设备。RF线圈,网络和前置放大器产生的热噪声水平的降低导致以3的顺序增强信噪比,同时将样品保持在其原始组成和自然线宽度中。该进度报告显示了我们的第一个600 MHz 3.2 mM HCN三重共振CPMAS CRYOPOBE原型获得的结果。

The probe is designed for standard bore magnets and compatible with the well proven CryoPlatform. Cryogenic cooling of the RF coil and preamplifier electronics yields a sensitivity gain of more than 3 compared to conventional probes. This gain is achieved with the sample staying at room temperature.

该探针的MAS速率为20 kHz,带有3.2 mm转子和MAS3单元。在寒冷状态下,可以通过新的探针提升,可以轻松且安全的转子交换,在寒冷状态下,可以降低CPMAS冷冻探针。自动调整和匹配增加了探针的舒适安全操作。

Technical Details

•¹³C and ¹⁵N –optimized triple resonance NMR probe

•增强¹和N的敏感性> 3倍

•One order of magnitude faster data acquisition and significantly increased productivity

•Simultaneous irradiation on ¹³C and ¹⁵N under strong proton decoupling

•非常适合对具有低标记水平和小分子的生物固体研究

•Automatic tuning, matching and magic angle adjustment and lift-assisted sample exchange

•样品温度范围:-20 C至+60 C

•MAS rates up to 20 kHz with specially designed 3.2 mm rotors

•Standard bore design, compatible with SB and WB magnets.

Applications

生物固体冷冻探针可以在生理温度下研究挑战性生物系统,而不会改变其样品组成。

This feature is extremely important in structural biology, as the integrity and temperature of the protein surrounding, are often crucial to maintain the biological function.

Diluted samples, such as labelled proteins in their native environment or small molecules in large assemblies, and samples in natural abundance or with low level of labelling, greatly benefit from the significant boost in sensitivity of this innovative probe.

在药理环境中,此探针允许扩展NMR工具箱。2d见C³C相关实验通常用于自然丰度中活性药物成分的高级表征。其他应用领域包括药物多态性和药物递送系统

更快的数据获取

淀粉样蛋白原和prions

~ 96.7 mg (u- ¹³C, ¹⁵N) HET-s (218-289) prion domain, courtesy of prof. A.Loquet, CNRS, Bordeaux, France.

由于光谱的增强敏感性recorded one order of magnitude faster and it’s possible to record the spectra with just a single scan and with an amazing signal-to-noise and resolution.

更快的数据获取
淀粉样蛋白原和prions

~ 96.7 mg (u- ¹³C, ¹⁵N) HET-s (218-289) prion domain, courtesy of prof. A.Loquet, CNRS Bordeaux, France.

由于光谱的增强敏感性recorded one order of magnitude faster and it’s possible to record the spectra with just a single scan and with an amazing signal-to-noise and resolution.

This results into a large time saving for the setup and acquisition of multidimensional experiments.

获得一个维度
淀粉样蛋白原和prions

〜96.7毫克(U- 13C,15N)HET-S(218-289)Prion域,由教授提供。A.Loquet,CNRS Bordeaux,法国。

通过Biosoldis冷冻探针提供的灵敏度,扫描的数量可以大大减少。结果,多维实验变得更快。然后可以在通常需要运行2D实验的同一时间执行3D实验。另外,4D实验,由于收购时间太长而通常不会进行,因此可以访问,需要少于一周的实验时间。

Explore Low Sensitivity Samples
Large biological assemblies

~ 7.5 mg of U – ¹³C, ¹⁵N – labeled Kif5b, a 349-residue protein, complexed with microtubules. Total sample amount ~ 80 mg, courtesy of prof. T.Pollenova, U.Delaware, USA.

Thanks to the sensitivity enhancement more challenging samples, more complex or less labeled, become the focus of the researcher.

We have recorded 3D NCACX and NCOCX experiments have been recorded on sample constituted of a labelled protein of about 350 residues in a complex with an unlabelled protein (total The total amount of isotopically labeled material is ~9 %).
These experiments, which lasted for ~5 and 6 day respectively, demonstrate the stability of the probe over several days of accumulation under multiple steps of simultaneous irradiation on all RF channels.

生物样品在其本地环境中
Bacteria cell envelope of mycobacteria

~ 40 mg ¹³C, ¹⁵N-labeled cell wall, HEPES 50 mM pH 7.0, courtesy of prof. J.P.Simorre, U.Grenoble, France.

Thanks to the sensitivity enhancement, the researchers can look with solid state NMR directly into cells.

This probe is a game changer for unstable samples that tends to degrade fast over time. On these kind of samples is crucial to be able to collect all the data shortly after they production.

Natural Products at Natural Abundance
Wood and Cellulose

Wood from Bordeaux wine threes, courtesy of prof. A.Loquet, CNRS Bordeaux, France.

Wood and other cellulose-rich samples typically present a mixture of amorphous and crystalline materials, and therefore a mixture of large and narrow signals. Distinguish between the different overlapping spectral components in a ¹D ¹³C experiment is often not easy. Being able to improve the resolution with a ²D CC correlation experiment is sometimes the only viable way to assign the spectral components. With the improved sensitivity of the BioSolids CryoProbe a refocused INADEQUATE become a routine tool.

Natural Products at Natural Abundance
本地胶原蛋白内部骨基质

Cortical femora bone of Goat (Capra hircus, 2-3 years old). N.Sinha, Centre of Biomedical Research, India.

了解胶原蛋白在健康和患病状况下胶原蛋白在其真正的本地状态下的结构和功能行为是开发各种骨变性疾病,组织工程和骨植入物治疗的里程碑。胶原蛋白的自然丰度光谱通常具有低灵敏度和有限的同位素浓度。
得益于该探针的灵敏度增强,现在可以记录自然同位素丰度中胶原蛋白的高分辨率2d 2d h-¹h-³c。这些光谱在短时间内的接触时间记录下来,阐明了骨基质内部在其真实天然环境中胶原蛋白的短距离相互作用。通过此探测,研究人员在理解骨基质内胶原蛋白的结构稳定性和功能机理方面迈出了一步。

In-situ Characterization of Active Pharmaceutical Ingredients
基于环糊精的金属有机框架(CD-MOF)

由教授提供的CDMOFS颗粒含有〜20%兰索拉唑。C.Martineau-Corcos,Cemhti UPR CNRS 3079,ILV UMR CNRS 8180和法国IUF。

Drug-delivery matrixes are often porous materials which resemble sponges. Metal-organic frameworks (CD-MOFs) are good example, as they are materials with a huge empty volume. Drugs loaded into MOFs represent a case of diluted spin systems, in which the sensitivity lack represent a big issue for the in-situ characterization of the active ingredient.

It is worth to note that the cryoprobe technology is so far the only tool that allows probing the state of a low API content in these kind of samples. Other approaches, like fast MAS 1H NMR or DNP have been demonstrated to be not suitable or of limited use, because of lack of resolution and unstability of CD-MOFs in the solvents used for DNP sample preparation.

A) Vitamin B12 B) 2D 13C-13C INADEQUATE on natural abundance Vitamin B12. 80 increments of 1024 scans each, acquired within a total experimental time of 3 days and 9 hours. The spectrum was processed using covariance.

Advanced Characterization of API

得益于探针的出色灵敏度,可以执行API的高级表征,而无需求助于同位素标记。在自然丰度下维生素B12上的CC双重量子相关性来说明这一点。重新聚焦不充分的敏感性低下,并且在RT探测器上将花费超过一个月的收购时间。

13 c和g的标准信噪比较lycine, illustrating the more than 3-fold enhancement factor compared to a traditional room temperature probe with the same amount of sample material. The spectrum shows that the magic angle is well set and that 1H high power decoupling is stable.

CP和基于双CP的实验

CPMAS冷冻探针是针对结构生物学的标准和高级基于CP的实验开发的。

将高灵敏度与NUS结合起来可用于显着改善光谱分辨率或减少实验时间。a)使用传统采样(〜4 h),在均匀标记的FMLF上显示CC相关性,使用25%NUS采样(〜1 h)显示了相同的实验。

将高灵敏度与NUS相结合

The high sensitivity of the new probe in combination with Non-Uniform Sampling techniques enables a dramatic reduction of experiment times. The CPMAS CryoProbe allows up to 10 times faster data acquisition for sensitivity limited samples.

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实验室

Service & Life Cycle Support for Magnetic Resonance and Preclinical Imaging

布鲁克’s commitment to provide customers with unparalleled help throughout the buying cycle, from initial inquiry to evaluation, installation, and the lifetime of the instrument is now characterized by the实验室service concept.

实验室Maintenance Agreements, On-Site On-Demand and Enhance Your Lab are designed to offer a new approach to maintenance and service for the modern laboratory

Publications

1。NPM1exhibitsstructuralanddynamicheterogeneityupon阶段separationwiththeP14arftumorsuppressor
Eric Gibbs, Barbara Perrone, Alia Hassan, Rainer Kümmerle, Richard Kriwacki.
磁共振杂志,2020,310,10646

2。CPMAS的灵敏度提高CryoProbefor Challenging Biological Assemblies
Alia Hassan, Caitlin M.Quinn, Jochem Struppe, Ivan V.Sergeyev, Chunting Zhang, Changmiao Guo, Brent Runge, Theint Theint, Hanh H.Dao, Christopher P. Jaroniec, Mélanie Berbon, Alons Lends, Birgit Habenstein, Antoine Loquet, Rainer Kuemmerle, Barbara Perrone, Angela Gronenborn, Tatyana Polenova.
Journal of Magnetic Resonance, 2020, 311, 106680

3.Efficient incorporation and protection of lansoprazole in cyclodextrin metal-organic frameworks
Xue Li, Marianna Porcino, Charlotte Martineau Corcos, Tao Guo, Ting Xiong, Weifen Zhu, Gilles Patriarch, Christine Péchoux, Barbara Perrone, Alia Hassan, Rainer Kümmerle, Alexandre Michelet, Anne Zehnacker-Rentien, Jiwen Zhang Ruxandra Gref.
International Journal of Pharmaceutics, 2020, 585, 119442

4.生物固体在骨基质内部的天然胶原蛋白中的短期和远程相互作用。
Nidhi Tiwari, Sebastian Wegner, Alia Hassan, Navneet Dwivedi, RamaNand Rai, and Neeraj Sinha.

在化学中接受的磁共振