Cryoprobe for 800 MHz NMR Spectrometer

冷冻探针技术的最新发展使溶液态NMR光谱仪的固有灵敏度大大提高。这一成就对于结构生物学领域的NMR研究具有巨大价值，因为仪器灵敏度在确定大多数生物分子NMR研究建议的可行性方面起着重要作用。目前的冷冻探针技术允许实现2至4倍范围内的灵敏度提高，确切的增益取决于样品缓冲液的电导率。相比之下，常规NMR探针的灵敏度仅提高20%是非常难以实现的，并且将被认为是相当显著的。冷冻探针的使用允许样品浓度降低2-4倍，这极大地促进了对具有低溶解度、聚集倾向或在样品材料特别难以获得和/或获得成本高的情况下的样品的工作。可替代地，通过使用冷冻探针，可以将总数据采集时间减少4至16倍，这在对稳定性有限的样品进行工作时，在考虑需要收集大量数据的项目时，对于允许在现实的时间段内执行低灵敏度实验的可能性，以及对于显著改善生物相容性，是非常有价值的。 非常昂贵的超高场NMR光谱仪的总吞吐量。为了使结构生物学家能够继续解决日益困难的问题，利用现有的任何技术进步至关重要。目前的提案要求提供资金，用于购买三重共振5 mm 1 H-{13 C，15 N}脉冲场梯度冷冻探针和800 MHz NMR光谱仪所需的附件，该光谱仪位于辛辛那提大学医学院的结构生物学/NMR设施内。该设施为大量NIH资助的研究人员提供支持，冷冻探针的获得将大大促进现有的研究项目，并将允许解决新的甚至更具挑战性的问题。

The recent development of cryoprobe technology is providing tremendous gains in the inherent sensitivity of solution-state NMR spectrometers. This achievement is of enormous value for NMR studies in the field of structural biology, due to the fundamental role of instrument sensitivity in determining the feasibility of most biomolecular NMR research proposals. Current cryoprobe technology has allowed sensitivity improvements by factors in the range of 2 to 4 to be realized, with the exact gain depending on the conductivity of the sample buffer. In contrast, improvements in sensitivity of conventional NMR probes by as little as 20% have been exceptionally difficult to achieve, and would be considered quite significant. Utilization of a cryoprobe allows sample concentrations to be reduced by factors of 2-4, which greatly facilitates work on samples with low solubility, a tendency to aggregate, or in cases where the sample material is especially difficult and/or costly to obtain. Alternatively, it is possible, by using a cryoprobe, to reduce total data acquisition times by factors of 4 to 16, which is extremely valuable when working on samples of limited stability, when considering projects which require extensive amounts of data to be collected, for allowing for the possibility of performing low-sensitivity experiments in realistic periods of time, and for significantly improving the overall throughput on very costly, ultra-high field NMR spectrometers. In order for structural biologists to continue to tackle increasingly difficult problems, it is vital to exploit whatever technological advances are available. The current proposal requests funds to purchase a triple resonance 5 mm1H-{13C,15N} pulsed field gradient cryoprobe and required accessories for the 800 MHz NMR spectrometer that is housed in the structural biology/NMR facility within the University of Cincinnati College of Medicine. This facility supports a large number of NIH-funded investigators, and acquisition of the cryoprobe will greatly facilitate the existing research projects and will allow new and even more challenging problems to be tackled.