Methods for protein-sds complex decoupling for protein mass spectrometry analysis

A method for offline protein-SDS complex decoupling for protein mass spectrometry analysis, comprising:

• obtaining a sample of a protein comprised in a buffer solution comprising sodium dodecyl sulfate (SDS);
• adding an amphipathic co-solvent to the buffer solution;
• adding a co-reagent to the buffer solution after the addition of the amphipathic co-solvent, wherein at least one of the amphipathic co-solvent and the co-reagent are pre-mixed with a protein stabilizer different than the amphipathic co-solvent, and wherein the additions of the amphipathic organic co-solvent and the co-reagent cause a decoupling of the SDS from at least a portion of the protein in the sample;
• and
• analyzing the decoupled protein using mass spectrometry.

A method for inline protein-SDS complex decoupling for capillary electrophoresis-mass spectrometry analysis, comprising:

• obtaining a sample of a protein comprised in a buffer solution comprising sodium dodecyl sulfate (SDS);
• preparing decoupling reagents comprising an amphipathic co-solvent, a protein stabilizer different than the amphipathic co-solvent, and a co-reagent;
• loading a capillary of a capillary electrophoresis-mass spectrometry (CE-MS) system with a background electrolyte;
• injecting the protein sample into the capillary;
• injecting the de-coupling reagents into the capillary, wherein the injection of the de-coupling reagents causes a decoupling of the SDS from at least a portion of the protein in the sample;
• and
• applying voltage in a positive or negative polarity to the capillary to start separation of the decoupled protein from the SDS via capillary electrophoresis, wherein the separated protein is analyzed via a mass spectrometer of the CE-MS system.