Online Hydrophobic Interaction Chromatography-Mass Spectrometry for the Analysis of Intact Monoclonal Antibodies.
Bifan ChenZiqing LinAndrew J AlpertCexiong FuQunying ZhangWayne A PrittsYing GePublished in: Analytical chemistry (2018)
Therapeutic monoclonal antibodies (mAbs) are an important class of drugs for a wide spectrum of human diseases. Liquid chromatography (LC) coupled to mass spectrometry (MS) is one of the techniques in the forefront for comprehensive characterization of analytical attributes of mAbs. Among various protein chromatography modes, hydrophobic interaction chromatography (HIC) is a popular offline nondenaturing separation technique utilized to purify and analyze mAbs, typically with the use of non-MS-compatible mobile phases. Herein we demonstrate for the first time, the application of direct HIC-MS and HIC-tandem MS (MS/MS) with electron capture dissociation (ECD) for analyzing intact mAbs on quadrupole-time-of-flight (Q-TOF) and Fourier transform ion cyclotron resonance (FTICR) mass spectrometers, respectively. Our method allows for rapid determination of relative hydrophobicity, intact masses, and glycosylation profiles of mAbs as well as sequence and structural characterization of the complementarity-determining regions in an online configuration.
Keyphrases
- mass spectrometry
- liquid chromatography
- tandem mass spectrometry
- high resolution mass spectrometry
- solid phase extraction
- high performance liquid chromatography
- ultra high performance liquid chromatography
- gas chromatography
- ms ms
- simultaneous determination
- capillary electrophoresis
- high resolution
- liquid chromatography tandem mass spectrometry
- endothelial cells
- ionic liquid
- social media
- molecularly imprinted
- magnetic resonance
- magnetic resonance imaging
- healthcare
- induced pluripotent stem cells
- computed tomography
- amino acid
- drug induced
- sensitive detection
- small molecule
- quantum dots
- protein protein