A. Carl Sanchez

The utility of native high-resolution mass spectrometry (HRMS) in intact protein characterization is rapidly growing because of advances in both ion-exchange chromatography (IEC) as well as MS-compatible buffer systems. MS is a critical component of biotherapeutic characterization, but its combination with traditional chromatographic separations, such as size-exclusion chromatography (SEC) and IEC, has been slow because of the predominant use of high salt mobile phases, which are incompatible with MS. Recently reported methods using cation-exchange chromatography (CEX) with volatile buffer systems for pH gradient elution has given researchers the ability to use these chromatographic techniques with MS detection. In this article a robust, MS-compatible buffer system for high sensitivity IEC with pH gradient elution for charge variant analysis of intact monoclonal antibodies (mAbs) is described.

A structured, general purpose approach to method development for bioanalytical hydrophilic interaction chromatography–tandem mass spectrometry (HILIC–MS-MS) applications is described.

The analysis of polar compounds in support of clinical and pre-clinical pharmacokinetic studies requires an analytical methodology capable of achieving ultra-low detection and quantification limits. The high sensitivity afforded by coupling HPLC with tandem mass spectrometry (MS–MS) has made it the technique of choice in this environment, but it is subject to the following limitations when reversed phase liquid chromatography (RPLC) is used