Automated Plasma Glycomics with Linkage-Specific Sialic Acid Esterification and Ultrahigh Resolution MS.
Gerda C M VreekerSimone NicolardiMarco R BladergroenCorné J van der PlasWilma E MeskerRob A E M TollenaarYuri E M van der BurgtManfred WuhrerPublished in: Analytical chemistry (2018)
High-throughput mass spectrometry (MS) glycomics is an emerging field driven by technological advancements including sample preparation and data processing. Previously, we reported an automated protocol for the analysis of N-glycans released from plasma proteins that included sialic acid derivatization with linkage-specificity, namely, ethylation of α2,6-linked sialic acid residues and lactone formation of α2,3-linked sialic acids. In the current study, each step in this protocol was further optimized. Method improvements included minimizing the extent of side-reaction during derivatization, an adjusted glycan purification strategy and mass analysis of the released N-glycans by ultrahigh resolution matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance MS. The latter resolved peak overlap and simplified spectral alignment due to high mass measurement precision. Moreover, this resulted in more confident glycan assignments and improved signal-to-noise for low-abundant species. The performance of the protocol renders high-throughput applications feasible in the exciting field of clinical glycomics.
Keyphrases
- mass spectrometry
- high throughput
- cell surface
- ms ms
- liquid chromatography
- high performance liquid chromatography
- gas chromatography
- randomized controlled trial
- multiple sclerosis
- liquid chromatography tandem mass spectrometry
- tandem mass spectrometry
- high resolution mass spectrometry
- simultaneous determination
- high resolution
- single molecule
- genome wide
- gas chromatography mass spectrometry
- single cell
- machine learning
- capillary electrophoresis
- optical coherence tomography
- air pollution
- solid phase extraction
- ultra high performance liquid chromatography
- hiv testing
- magnetic resonance imaging
- electronic health record
- magnetic resonance
- computed tomography
- molecularly imprinted
- high density
- men who have sex with men