4D中的OMICS
蒂姆斯添加了分离的第四维
基于质谱(MS)的蛋白质组学已成为鉴定和量化数千种蛋白质的强大技术。但是,由于当前质谱仪的速度,灵敏度和分辨率有限,完整蛋白质组织的覆盖范围仍然非常具有挑战性。
The timsTOF Pro uses the Parallel Accumulation Serial Fragmentation (PASEF®) acquisition method to provide extremely high speed and sensitivity to reach new depths in shotgun proteomics and phosphoproteomics, using low sample amounts.
Trapped Ion Mobility Spectrometry (TIMS) is first and foremost a separation technique in gas phase, which resolves sample complexity with an added dimension of separation in addition to HPLC and mass spectrometry, increasing peak capacity and confidence in compound characterization. Equally as important, the TIMS device also serves to accumulate and concentrate ions of a given mass and mobility, enabling a unique increase in sensitivity and speed along with the additional dimension of separation.
实现shot弹枪蛋白质组学的接近100%占空比
对于高灵敏度,现在可以通过双重TIMS技术实现高速shot弹枪蛋白质组学的接近100%占空比。新颖的设计使离子积聚在前部,而后部的离子则根据其离子迁移率依次释放。此过程称为平行累积串行片段或Pasef®。
第二代双TIMS分析仪针对shot弹枪蛋白质组学需求进行了优化。由于其独特的几何离子,释放了从TIMS分析仪的第二部分中的移动性释放,而进一步的离子可以在TIMS分析仪的第一部分并行积累。使用平行的积累技术,获得接近100%的占空比,导致几乎没有离子损失。
In the timsTOF Pro an advanced segmented quadrupole mass filter is used for high ion transmission and isolation efficiency. The quadrupole mass position is synchronized with the elution times of the specific ions from the TIMS analyzer. Due to its ultra-high mass position switching time (< 1 ms) it enables the best performance for the PASEF®method.
With speed in mind, Bruker experts redesigned MS/MS technology to meet the requirements of shotgun proteomics. Peptide ions are separated using trapped ion mobility spectrometry, eluted (~ 100 ms) and detected in the QTOF, generating the TIMS MS heat map. In the PASEF®方法在洗脱过程中,将某个离子物种隔离并立即转移到下一个前体。父和片段光谱由迁移率值对齐。与平行的积累串行碎片(Pasef)®)技术> 100 Hz测序速度。与pasef®方法可以通过选择多次来增加低丰富肽的MS/MS光谱质量。
Pasef®:非常适合shot弹枪蛋白质组学
The timsTOF Pro powered by PASEF®提供测序速度> 100 Hz而不会失去灵敏度或分辨率。这是通过将四极隔离质量窗口与来自TIMS漏斗的特定肽包装的洗脱时间同步的。
The only PASEF®-enabled mass spectrometer
布鲁克(Bruker)在2016年推出了TIMS(被困的离子移动光谱法)技术,作为一种革命性的离子移动技术,在极其紧凑的设备中实现了前所未有的离子迁移率分辨率。第二代双TIMS分析仪现在启用PASEF®(平行积累串行碎片)正在为shot弹枪蛋白质组学设定不妥协的速度,灵敏度和分辨率的新标准。
Superior robustness, no cleaning required
许多用于shot弹枪蛋白质组学的MS仪器需要每两周或每月清洁,每天在大型样品队列上运行24小时,并且在较短的时间段内,性能降解值得注意。Timstof Pro的出色鲁棒性意味着可以在多天或几周内24/7运行该仪器,而不会明显丧失灵敏度或其他性能指标。
TIMSTOF PRO允许在超过100 Hz的测序速度下分析数百个具有最小样品负载(<200 ng)的样品。所有这些都具有毫不妥协的蛋白质组学深度。这改变了蛋白质组学的完成方式,但也释放了新的约束数据分析。现代研究人员必须使用Timstof Pro处理数百个信息密集内容样本。对于大多数管道,这在数据分析步骤中会产生瓶颈。Bruker通过介绍称为PASER的实时数据库搜索功能(实时数据库搜索引擎)来创新这一步骤,该功能可删除数据分析障碍。使用PASER,一旦完成LC-MS运行,结果就可以手动或运行。
最大/Perseus and PEAKS Studio data processing
An open-file data format allows researchers to work directly with the raw data and use industry leading software. MaxQuant has been adapted to manage 4-dimensional (4D) features in the space spanned by retention time, ion mobility,
mass, and signal intensity which benefit the identification and quantification of peptides, proteins, and posttranslational modifications. PEAKS Studio combines de novo sequencing with traditional database searches and is optimized for processing timsTOF raw data.
prm-PASEF®: enabling high-throughput, high sensitivity targeted proteomics
Enabling high-throughput, high sensitivity targeted proteomics. The prm-PASEF®acquisition method has been developed to translate the advantages of the parallel accumulation serial fragmentation (PASEF®)获取靶向蛋白质组学领域的获取策略。
Proteomic Interrogation of Primary Insulin-secreting Pancreatic β-cells
在Timstof Pro上的综合离子迁移率与经经速度的MS/MS采集的组合为对β细胞生理学的深度,分子水平的理解和糖尿病的潜在新目标途径铺平了道路。
高吞吐量4D-蛋白质组™ - DIA-PASEF的应用®和Evosep One用于短梯度
TimStof Pro提供了两种独特技术的组合,即由捕获的离子迁移率光谱法提供的第四维(TIMS),以增强离子分离和灵敏度和平行积累串行片段(PASEF(PASEF)®)
to improve ion utilization efficiency and data acquisition speed.
"We now know that the peptide mixtures are still extremely complex when analyzing them in two dimensions (retention time and m/z). Adding one more dimension should in principle get us a long way ahead. In addition to the additional dimension of separation, the timsTOF Pro gives us extremely high speed and sensitivity to get deeper into the proteome and using less sample material.”
For Research Use Only. Not for use in clinical diagnostic procedures.