冷冻探针

生物固体冷冻探针

提高灵敏度而没有样品修改

While being extremely versatile, NMR has the inherent challenge of being insensitive. One strategy to improve the sensitivity which does not require sample modification is cryogenically cooling the RF coil and preamplifier electronics. The reduced level of thermal noise generated by the RF coil, network, and preamplifier lead to a boost in signal-to-noise on the order of 3 while keeping the sample in its original composition and with its natural line width. This progress report shows results obtained with our first 600 MHz 3.2 mm HCN triple resonance CPMAS CryoProbe prototype.

该探针是为标准孔磁铁设计的，与经过验证的冷冻文化形式兼容。与常规探针相比，RF线圈和前置放大器电子产品的低温冷却超过3。样品保持在室温下，可以实现这一增益。

The probe reaches MAS rates of 20 kHz with 3.2 mm rotors and the MAS3 unit. The CPMAS CryoProbe can be lowered while in the cold state with a new probe lift allowing for easy and safe rotor exchange. Automatic tuning and matching adds to the probe’s comfortable and safe operation.

技术细节

•Å³C和n - 优化的三重共振NMR探针

•Enhancement ¹³C and ¹⁵N sensitivities by a factor of >3

•一个数量级更快的数据获取，并显着提高了生产率

•在强质子脱钩下同时辐照它们和⁵N

•Ideally suited for research on biosolids with low level of labeling and small molecules in natural abundance

•自动调整，匹配和魔术角度调整以及举重辅助样品交换

•Sample temperature range: -20 C to +60 C

•MAS速率高达20 kHz，专门设计的3.2毫米转子

•标准孔设计，与SB和WB磁铁兼容。

申请

The BioSolids CryoProbe allows to study challenging biological systems under physiological temperatures and without altering their sample composition.

由于蛋白质周围的蛋白质的完整性和温度通常对于维持生物学功能至关重要，因此此特征在结构生物学中非常重要。

稀释的样品，例如在其天然环境中标记的蛋白质或大型组合中的小分子，以及具有自然丰度或低标记水平的样品，从该创新探针的敏感性显着提高。

In the pharmacological context, this probe allows to expand the NMR toolbox. 2D ¹³C¹³C correlation experiments can be routinely used for advanced characterization of active pharmaceutical ingredients in natural abundance. Other fields of applications include drug polymorphism and drug-delivery systems

更快的数据采集

Amyloid fibrils and Prions

〜96.7毫克（u-κC，n）Het-S（218-289）Prion域，由教授提供。A.loquet，CNRS，波尔多，法国。

Thanks to the enhanced sensitivity the spectra are 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.

更快的数据采集
Amyloid fibrils and Prions

〜96.7毫克（u-κC，n）Het-S（218-289）Prion域，由教授提供。A.Loquet，CNRS Bordeaux，法国。

这导致了大量节省的设置和获取多维实验。

Gain One Dimension
Amyloid fibrils and Prions

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

With the sensitivity provided by the BioSoldis CryoProbe the number of scans can be drastically reduced. As a consequence, the multidimensional experiments become faster. It is then possible to perform a 3D experiment in the same time normally required to run a 2D experiment. Also 4D experiments, routinely not performed because of too long acquisition time, become accessible, requiring less than a week of experimental time.

探索低灵敏度样本
大型生物组装

〜7.5毫克的U - ¹³C，n - 标记为KIF5B，一种349个剩余蛋白，与微管复合。总样本量〜80 mg，由教授提供。美国U.Delaware的T.Pollenova。

由于灵敏度增强，更具挑战性的样本，更复杂或标记的样本成为研究人员的重点。

我们已经记录了3D NCACX，并且已经记录了NCOCX实验，该样品是由未标记蛋白的复合物中约350个残基的标记蛋白组成的（同位素标记材料的总数约为9％）。
这些实验分别持续了〜5和6天，证明了探针在所有RF通道上同时辐照的多个步骤下积累的稳定性。

Biological Samples in their Native Environment
分枝杆菌的细菌细胞信封

〜40毫克，卡，标记的细胞壁，Hepes 50 mm pH 7.0，由教授提供。J.P. Simorre，法国U.Grenoble。

得益于灵敏度的增强，研究人员可以将固态NMR直接介绍到细胞中。

该探测器是不稳定样本的游戏规则改变者，随着时间的流逝，往往会迅速降解。在这些样品上，对于能够在生产后不久收集所有数据至关重要。

天然产品的天然产品
木材和纤维素

来自波尔多葡萄酒三人的木材，由教授提供。A.Loquet，CNRS Bordeaux，法国。

木材和其他富含纤维素的样品通常呈现无定形和结晶材料的混合物，因此是大小信号的混合物。必威手机客户端区分不同的重叠光谱成分在实验中通常并不容易。有时能够通过二级CC相关实验改善分辨率是分配光谱成分的唯一可行方法。随着生物固体的敏感性的提高，冷冻螺旋桨的重新聚焦不足成为常规工具。

天然产品的天然产品
Native collagen inside bone matrix

山羊皮质股骨（Capra Hircus，2-3岁）。N.Sinha，印度生物医学研究中心。

Understanding the structural and functional behaviour of collagen in its true native state in healthy and diseased conditions is a milestone for the development of treatments for various bone degenerative diseases, tissue engineering, and bone implants. Natural abundance spectra of collagen typically suffer of low sensitivity and limited isotopic concentration.
Thanks to sensitivity enhancement of this probe, it is now possible to record high-resolution 2D ¹H-¹³C HETCOR experiments of collagen in natural isotopic abundance. The spectra, recorded at short and long contact times , shed light on short and long-range interactions of collagen inside the bone matrix in its true native environments. With this probe, the researchers go one step further in the understanding of the structural stability and functional mechanism of collagen inside the bone matrix.

活性药物成分的原位表征
Cyclodextrin-based metal-organic frameworks (CD-MOFs)

CDMOFs particles loaded with ~ 20 % Lansoprazole, courtesy of prof. C.Martineau-Corcos, CEMHTI UPR CNRS 3079, ILV UMR CNRS 8180, and IUF, France.

药物传递基质通常是类似于海绵的多孔材料。必威手机客户端金属有机框架（CD-MOF）就是一个很好的例子，因为它们是具有巨大空体积的材料。必威手机客户端加载到MOF中的药物代表了稀释的自旋系统的情况，其中缺乏灵敏度代表了活性成分的原位表征的一个大问题。

值得注意的是，迄今为止，冷冻探针技术是唯一允许在此类样品中探测低API含量状态的工具。由于缺乏用于DNP样品制备的溶剂中CD-MOF的分辨率和不稳定，因此已证明其他方法（例如快速MAS 1H NMR或DNP）不适合使用或有限使用。

a）维生素B12 b）2d 13c-13c自然丰度维生素B12不足。80个增量为1024次扫描，在3天和9小时的总实验时间内获取。使用协方差处理频谱。

API的高级表征

Thanks to the superior sensitivity of the probe, advanced characterization of an API can be performed without resorting to isotopic labeling. This is illustrated with a CC double quantum correlation on vitamin B12 at natural abundance. The refocused INADEQUATE has notoriously low sensitivity, and it would have taken more than one month acquisition time on a RT probe.

13C与甘氨酸的标准信噪比比较，与传统的室温探针相比，具有相同数量的样品材料的传统室温探针相比，增强因子超过3倍。频谱表明，魔术角度设置好，1H高功率解耦稳定。

CP and Double-CP based Experiments

The CPMAS CryoProbe is developed for standard and advanced CP-based experiments in structural biology.

Combining the high sensitivity with NUS can be used to significantly improve the spectral resolution or reduce the experimental time. A) shows a CC correlation on uniformly labeled fMLF using traditional sampling (~4 h), B) shows the same experiment using 25% NUS sampling (~1 h).

Combining High Sensitivity with NUS

新探针与非均匀抽样技术的高灵敏度可显着减少实验时间。CPMAS冷冻探针可为有限的样品允许更快的数据采集速度。

Service and Support

LabScape

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布鲁克致力于在整个购买周期内为客户提供无与伦比的帮助，从初始查询到评估，安装和仪器的寿命，现在以此为特征实验室服务概念。betway手机客户端下载

实验室维护协议，现场点播和增强实验室旨在为现代实验室提供一种新的维护和服务方法betway手机客户端下载

实验室小册子

出版物

1.NPM1 展览结构和动态的异质性之上 phase 分离和这 p14ARF 瘤抑制器
埃里克·吉布斯（Eric Gibbs），芭芭拉·佩伦（Barbara Perrone），阿里亚·哈桑（Alia Hassan），雷纳·库梅尔（RainerKümmerle），理查德·克里瓦奇（Richard Kriwacki）。
Journal of Magnetic Resonance, 2020, 310, 10646

2.Sensitivity Boosts by the CPMAS 冷冻探针用于挑战性的生物组装
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。
磁共振杂志，2020，311，106680

3。在环糊精金属有机框架中的有效掺入和保护兰索拉唑
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.
国际药品杂志，2020，585，119442

4.Probing short and long-range interactions in native collagen inside the bone matrix by BioSolids CryoProbe
Nidhi Tiwari，Sebastian Wegner，Alia Hassan，Navneet Dwivedi，Ramanand Rai和Neeraj Sinha。

Accepted in Magnetic Resonance in Chemistry