Automating Native Mass Spectrometry Through the Use of Online Buffer Exchange

Native mass spectrometry (nMS), in which intact protein or nucleoprotein complexes are directly measured, commonly involves exchange of nonvolatile electrolytes (NaCl, PBS, Tris) for volatile electrolytes such as ammonium acetate. This can be time-consuming if offline buffer exchange spin cartridges are used. Furthermore, some samples may not be stable if left in nonvolatile buffer for more than just a very short period of time. This problem can be avoided if an LC-compatible online buffer exchange column is used to separate protein complex from salt.

In this experiment, no attempt is made to perform size exclusion chromatography to separate protein complexes of different sizes but rather any protein complex that is injected onto the system is simply separated from low mass salts that would otherwise product heavily adducted protein complexes that appear as broad peaks in the mass spectrum.

This webinar will focus on the development of online buffer exchange (OBE) for nMS applications and will describe how the experiment is implemented and how it can be extended by coupling to affinity separation (e.g., IMAC-OBE) to be used to optimize protein overexpression.

Key Learning Objectives:

• The use of online buffer exchange for high throughput native mass spectrometry
• Online buffer exchange nMS allows to directly screen structural features of large biomolecules even if such biomolecules are not very stable in MS friendly buffers
• How the coupling of affinity separation (e.g., IMAC-OBE) to online-buffer exchange can be used to optimize protein overexpression

Who Should Attend:

• MS practitioners
• Researchers/ R&D Managers
• Laboratory Managers/ Directors / Supervisors
• Laboratory Technicians / Operators
• Structural Biologists
• Scientists in biopharma engaged in biotherapeutic characterization

Presenter: Vicki Wysocki, Ph.D. (Professor, Dept of Chemistry and Biochemistry, The Ohio State University)

Presenter: Catherine Dold (Health & Environment Writer, C&EN Media Group)