Protein drugs represent a challenge for separations by liquid chromatography (LC) because of the multiplicity of species and the inherent difficulty of separating large molecules with high resolution. Further, it is desirable to identify the proteins in each peak by mass spectrometry (MS), but there has always been a trade-off between LC resolution and MS sensitivity. In this research, we investigate polymer brushes as bonded phases to alleviate the trade-off in performance between LC and MS. The concept is that the polymer can be thicker than traditional silane bonded phases, thereby screening silanophilic interactions. This alleviates the need for trifluoroacetic acid, which would otherwise decrease MS sensitivity due to its high interfacial activity. The polymer brush is shown to allow for less acidic additives, such as formic acid, to allow for high sensitivity in MS, yet still provide for excellent resolution in LC. The results show that sensitive, high resolution LCMS is enabled for monoclonal antibody drugs, including IgG1, IgG2 and IgG4.

Polymer brushes as bonded phases for LCMS of protein drugs

255th National Meeting (2018)

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