Buffer Exchange Mass Spectrometry

Sample purity, integrity, and oligomeric state was analyzed by on-line buffer exchange MS in 200 mM ammonium acetate using a Vanquish ultra-high performance liquid chromatography system coupled to a Q Exactive ultra-high mass range Orbitrap mass spectrometer (Thermo Fisher Scientific). A self-packed buffer exchange column was used (P6 polyacrylamide gel, BioRad) (44 (link)). The recorded mass spectra were deconvolved with UniDec version 4.2+ (45 (link)).

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Sahtoe D.D., Praetorius F., Courbet A., Hsia Y., Wicky B.I., Edman N.I., Miller L.M., Timmermans B.J., Decarreau J., Morris H.M., Kang A., Bera A.K, & Baker D. (2022). Reconfigurable asymmetric protein assemblies through implicit negative design. Science (New York, N.Y.), 375(6578), eabj7662.

Publication 2022

Vanquish ultra-high performance liquid chromatography system Q Exactive ultra-high mass range Orbitrap mass spectrometer P6 polyacrylamide gel

Ammonium acetate Buffer High performance liquid chromatography Mass spectra Polyacrylamide gel

Corresponding Organization : Howard Hughes Medical Institute

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Variable analysis

independent variables

Buffer exchange column used (P6 polyacrylamide gel, BioRad)

dependent variables

Sample purity
Sample integrity
Sample oligomeric state

control variables

Buffer: 200 mM ammonium acetate
Instrumentation: Vanquish ultra-high performance liquid chromatography system coupled to a Q Exactive ultra-high mass range Orbitrap mass spectrometer (Thermo Fisher Scientific)

controls

No positive or negative controls were explicitly mentioned in the input.

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