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Rapid Online Buffer Exchange: A Method for Screening of Proteins, Protein Complexes, and Cell Lysates by Native Mass Spectrometry

preprint
revised on 16.09.2019 and posted on 17.09.2019 by Zachary VanAernum, Florian Busch, Benjamin J. Jones, Mengxuan Jia, Zibo Chen, Scott E. Boyken, Aniruddha Sahasrabuddhe, David Baker, Vicki Wysocki
It is important to assess the identity and purity of proteins and protein complexes during and after protein purification to ensure that samples are of sufficient quality for further biochemical and structural characterization, as well as for use in consumer products, chemical processes, and therapeutics. Native mass spectrometry (nMS) has become an important tool in protein analysis due to its ability to retain non-covalent interactions during measurements, making it possible to obtain protein structural information with high sensitivity and at high speed. Interferences from the presence of non-volatiles are typically alleviated by offline buffer exchange, which is timeconsuming and difficult to automate. We provide a protocol for rapid online buffer exchange (OBE) nMS to directly screen structural features of pre-purified proteins, protein complexes, or clarified cell lysates. Information obtained by OBE nMS can be used for fast (<5 min) quality control and can further guide protein expression and purification optimization.

Funding

P41 GM128577

S10 OD018507

History

Email Address of Submitting Author

vanaernum.2@osu.edu

Institution

The Ohio State University

Country

United States

ORCID For Submitting Author

0000-0002-0956-3956

Declaration of Conflict of Interest

The authors declare that they have no competing financial interests.

Version Notes

V2: added figures in line with text. V3: Minor grammatical fixes. V4: Updated manuscript with additional figures.

Exports