How to run a high-mass CCS calibration - WKB121398

OBJECTIVE or GOAL

Run a CCS calibration above 2000 Da.

ENVIRONMENT

• SYNAPT G2
• SYNAPT G2 MS/HDMS
• SYNAPT G2-S MS/HDMS
• SYNAPT G2-Si MS/HDMS
• SYNAPT G2-S
• SYNAPT G2-Si
• SYNAPT XS
• MALDI SYNAPT G2-Si
• Vion IMS QTof

PROCEDURE

If you want to calibrate CCS in higher mass ranges, you must find suitable calibrant molecules, substances with known CCS values typically available from the online repositories of academics such as David Clemmer, John MacLean, and Matthew Bush. In this range it is native protein peaks of lower charge states in neutral pH (holomyoglobin, ADH, concanavalin, etc.). You can pick the reference values based on what you can obtain for the acquisition from Sigma or other sources. You would then adjust the IMS conditions to transmit these heavy molecules and run typically a nanoESI offline native spray MS acquisition in IMS mode. For applying the calibration, you can use Driftscope 2.9. In the attached PDF document, the procedure starts at about page 50.

ADDITIONAL INFORMATION

The 5000 mass range requires manual calibration using Driftscope. We recommend the CCS references from the Bush lab:
https://depts.washington.edu/bushlab/ccsdatabase/