Targeted, hypothesis driven protein/peptide quantitation using MRM enables you to detect and measure hundreds of proteins in a given experiment at high sensitivity. The AB SCIEX Triple Quad and QTRAP LC/MS/MS Systems are the industry's premier systems for MRM based peptide and protein quantitation.
Highest precision MRM assays at the lowest levels of quantitation
- Fast and easy assays - no ELISA's or antibodies required
- High reproducibility with %CV typically less than 5%
- Measure hundreds of proteins in a single run, with multiple peptide measurements for each protein using the Scheduled MRM™ algorithm
- Ultimate confidence in the results using MS/MS confirmation of MRM assays in the same experiment
Fast, robust, high sensitivity assay development without protein/peptide standards
Taking advantage of unique TripleTrap™ scanning that switches the QTRAP® system between triple quadrupole and linear ion trap mode in under 1 millisecond, peptide MRM assays can be developed quickly and cheaply.
- QTRAP technology delivers up to 100x sensitivity in MS/MS for development of quantitative MRM assays for lower level proteins and peptides.
- MRMPilot™ Software automates the development of these optimized MRM assays.
- Quickly develop assays to many candidate markers directly from the discovery sample
- Create specific assays for protein isoforms
Unique and Powerful MRM3 Workflow for Quantitation of Peaks with Interferences
There are times when good quantitative results can't be achieved with a specific peptide SRM/MRM transition due to high background or interferences. Often the solution is to simply switch to another peptide or different fragment ion to obtain better results. However, there may be cases where this isn't practical, such as when a specific peptide or transition must be monitored due to tracking a specific protein isoform or modification. The MRM3 experiment on the QTRAP® 5500 system is a unique scan-type specifically designed to help achieve better quantitative results for those MRM transitions that are suffering from high background or interferences. Because MRM3relies on the quantitation of a second generation fragment ion, this additional filtering step enables a much cleaner background. As an added benefit, the use of MRM3 in place of MRM can help to relax the constraints around extensive sample clean-up.
Fig 1: Quantitation of a 39 amino acid peptide (MW 4186.6 Da) in plasma. Top: MRM showing strong interfering shoulder to the right of the peptide peak. Injection amount was 5 ng/mL on column. Bottom: The same amount injected using an MRM3experiment, now showing clean background. The use of MRM3 allowed >20x improvement in LOQ vs. MRM.
|
AB SCIEX QTRAP® 5500 System for Targeted Protein and Peptide Quantitation
The most sensitive and accurate instrument for peptide quantitation
The QTRAP® 5500 System combines a high sensitivity Triple Quadrupole with a fast and extremely sensitivity Linear AcceleratorTM Trap for high performance peptide quantitation. During assay development, this combination enables the development of MRM/SRM assays quickly and efficiently without the use of peptide standards using the MIDASTM workflow - a unique scan-type only available on QTRAP® based systems that combines MRM quantitation with full scan, high sensitivity MS/MS confirmation. The full scan MS/MS data is invaluable for optimizing and troubleshooting MRM transitions and makes assay design much faster and ultimately more robust.
- Highest sensitivity MRM instrument for peptide quantitation
- Highest sensitivity Linear AcceleratorTM Trap for simultaneous full scan MS/MS confirmation
- LINAC® Collision cell enables thousands of MRM transitions without cross-talk
- AcQuRateTM Pulse Counting Detector for accurately detecting single-ion events - extremely important for quantifying low level analytes
|
Fig 1: Assay Development using the MIDASTM Workflow. Top Left: XIC of a peptide MRM transition for coagulation factor XII in depleted plasma showing two major peaks. Bottom Left: Examining the full scan MS/MS spectrum under each peak indicates that the second peak is for coagulation factor (full scan MS/MS data matches theoretical sequence for coagulation factor peptide). The predicted MRM was the Y4 ion above the parent ion, however, it is clear from the fragmentation pattern that the y7 ion might be a better Q3 candidate. Right: Using the y7 ion for the MRM assay improves the S/N by > 10x. (Note: a longer chromatographic run was used for the initial detection and then the speed and sensitivity were simultaneously improved with help from the MIDAS Workflow for subsequent optimizations and the creation of the final assay).
+ View Larger
|
|
One combination of offerings to help you perform targeted peptide and protein quantitation analysis is below:
Instruments
|
 |
AB SCIEX QTRAP® 5500 LC/MS/MS system: This next-generation technology is designed to excel at peptide quantitation as a result of uniting the world's most sensitive triple quadrupole with innovative Linear AcceleratorTM trap technology. Intelligent linking of the system's capabilities with TripleTrapTM scanning enables powerful and unique quantitative and qualitative experiments such as the MIDASTM workflow. |
|
 |
AB SCIEX TripleTOFTM 5600 System: The AB SCIEX TripleTOFTM 5600 System is the only Accurate mass instrument with the speed and sensitivity of a triple quadrupole providing users with the ability to achieve high sensitivity quantification and advanced qualitative capabilities for rapid protein / peptide identification and preliminary peptide verification experiments on a single platform. |
Software
|
 |
MRMPilotTM Software: This software streamlines the building and optimization of targeted peptide quantitative assays on AB SCIEX QTRAP®, TripleTOFTM, and Triple QuadTM systems for biomarker verification or other targeted quantitative proteomic applications. |
|
 |
MultiQuantTM Software: MuliQuant Software is a powerful and easy-to-use software quantitation package that processes MRM & TOF MS data for quantitative information. The software easily handles large data sets consisting of both large numbers of MRM transitions and study samples. Results can be easily exported to other software packages or use the software's flexible reporting features to generate custom reports. |
|
View brochures, technical notes, webinars and videos about how we can help you quantitate proteins and peptides.
Media
Brochures
Technical Notes
Video
|
