GEN-MKT-18-7897-A
Sep 15, 2015 | Blogs, Life Science Research, Proteomics | 0 comments
Frankly, who has the time? With so many samples to prepare, day after day, week after week, month after month, there’s time for little else. And those repetitive tasks of pipetting over and over again, just keep, well, repeating.
It’s time to automate your sample prep! In this technical note, see how researchers reduce day-to-day variability resulting from manual, time-consuming, and multi-step protocols such as the protein denaturation, reduction, alkylation and digestion steps which precede LC/MS analysis. Here, the researchers use a Biomek NXP Span-8 Workstation and protein preparation kits with ready-to-use reagents to automate the entire workflow. Then, they tested the automated workflow for their quantitative proteomics studies.
First, the reproducibility of the digestion protocol alone was tested. As shown in the technical note, very high reproducibility was obtained, with about 90% of peptides monitored by LC/MS having digestion reproducibility below 10% CVs. Next, the digestion protocol was transferred to the Biomek NXP Span-8 Workstation. Many of the proteotypic peptides analyzed showed very high reproducibility with %CVs ~ 2-3%. As expected, for both the manual and automated protocol some peptides showed higher variability than others from the same protein, reflecting inherent digestion heterogeneity. When developing assays for peptide/protein quantitation, it’s important to assess the variability in digestion reproducibility in order to choose the highest performing peptides. Automation of this step can greatly simplify the assessment.
Finally, the researchers tested the reproducibility across different labs and workstations, and across multiple days. The results show very similar cumulative reproducibility curves with more than 80% of peptides monitored having CVs below 10%, demonstrating that the automation method and protocol were very robust and transferable.
To learn more, download the full technical note and see how the Biomek Workstation with optimized protein preparation kits can help you save time while providing reproducible sample preparation today.
Now, what will you do with all that free time?
The Echo® MS+ system is a novel platform for Acoustic Ejection Mass Spectrometry (AEMS) and combines the speed of acoustic sampling with the selectivity of mass spectrometry. This platform has been designed for high throughput analysis of small and large molecules. The technology combines Acoustic Droplet Ejection (ADE), an Open Port Interface (OPI) and could be coupled with the SCIEX Triple Quad 6500+ system or the ZenoTOF 7600 system.
The Echo® MS+ system comprises of an open-port interface (OPI) and acoustic droplet ejection (ADE) module which could be coupled with a mass spectrometer. The mass spectrometer could either be a SCIEX Triple Quad 6500+ system or the ZenoTOF 7600 system. This non-liquid chromatography based; high-throughput screening platform enables rapid analysis of compounds at speeds of up to 1 sample/second.
The ability to consistently achieve reproducible results on many complex samples across multiple days is critical to a routine clinical laboratory. Laboratories relying on analytical instrumentation require stability and robustness to perform a variety of screening and confirmatory assays with confidence. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) has become the preferred analytical method in the clinical laboratory to reliably perform clinical testing as it provides best-in-class performance and reliability for the most challenging assays. LC-MS/MS offers the required levels of sensitivity and specificity for the detection and quantitation of molecules from complex biological samples, helping laboratories deliver highly accurate data for a variety of clinically relevant analytes across a wide range of assays.
Posted by
You must be logged in to post a comment.
Share this post with your network