Implementation of Infrared-Activated Negative Electron Transfer Dissociation (IR-NETD) Using Xenon on a...

Area of Science:

• Analytical Chemistry
• Mass Spectrometry
• Biochemistry

Background:

• Photoactivation in tandem mass spectrometry offers advanced dissociation strategies.
• Hybrid mass spectrometry systems require versatile dissociation techniques for complex molecule analysis.

Purpose of the Study:

• To implement an infrared (IR) laser system on a quadrupole-Orbitrap-quadrupole linear ion trap hybrid MS system.
• To establish xenon as an efficient radical cation source for negative electron transfer dissociation (NETD).
• To evaluate the performance of IR-activated NETD (IR-NETD) for characterizing RNA molecules, including complex small interfering RNA (siRNA).

Main Methods:

• Integration of a home-built photon detector and xenon introduction into a hybrid MS system.
• Assessment of IR-NETD using a 6-mer RNA molecule.
• Characterization of a complex siRNA molecule using IR-NETD, analyzing precursor charge state and IR laser power.

Main Results:

• Successful implementation of an IR laser system on the Orbitrap Ascend hybrid MS.
• Demonstration of xenon as an effective source for NETD reactions.
• Validation of IR-NETD for characterizing both simple and complex RNA molecules, highlighting the benefits of concurrent IR photoactivation.

Conclusions:

• The developed instrumental approach is powerful and versatile for biopharmaceutical molecule characterization.
• IR-NETD offers an efficient method for analyzing complex RNA structures.
• This technique enhances the capabilities of hybrid mass spectrometry for detailed molecular analysis.