Date: | 05/03/2017 |
Categories: | Academia Omics , Pharma CRO , Analyst Software |
For research use only. Not for use in diagnostic procedures.
Issue Description
Using the TripleTOF™ 5600 and 6600 Intact Protein Analysis Script
The Intact Protein Analysis script is used whenever proteins with MW > ~ 6kDa are analyzed on the 5600. The script makes modifications that improve the analysis of both small and large proteins and should be used in all cases.
THIS IS A PRE-BETA VERSION OF THE SCRIPT AND IS TO BE USED ONLY BY ABSCIEX PERSONNEL ONLY. IT IS NOT FOR CUSTOMER DISTRIBUTION.
Installation of the Script:
Resolution
How will I know it is working?
When analyzed using default settings on the 5600 proteins will show certain easily recognized artifacts. Smaller proteins for which isotopic resolution is possible will show a marked suppression of the low m/z peaks in the isotope cluster. This is shown below for horse myoglobin (5pmol injection) for a single scan spectrum near the apex of the LC peak:
With Intact Protein Mode activated the isotope cluster will not show this artifact and will have a Gaussian shape.
For larger proteins (MW> ~25 kDa) the use of default settings may result in splitting on the low m/z side of the peak as well as consistent dropping out of the signal in the baseline. This is shown below for bovine apo-transferrin (10 pmol injection) for a single scan spectrum near the maximum of the LC peak:
Activation of Intact Protein Mode will generally result in considerably improved signal-top-noise for the protein peaks, as well as elimination of the peak splitting and signal dropouts:
Time Bins to Sum:
For proteins for which isotopic resolution is desired (MW<20 kDa) it is recommended that a Time Bins to Sum value of 1 be selected. For larger proteins a value of 10 is recommended.
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