Dataset Information

Higher Temperature Porous Graphitic Carbon Separations Differentially Impact Distinct Glycopeptide Classes

ABSTRACT: Mass spectrometry-based discovery glycoproteomics is highly dependent on the use of chromatography paradigms amenable to analyte retention separation. When compared against established stationary phases such as reversed phase and hydrophilic interaction liquid chromatography, reports utilizing porous graphitic carbon (PGC) have detailed its numerous advantages. Recent efforts have detailed the utility in porous graphitic carbon in high throughput glycoproteomics, principally through enhanced profiling depth and liquid phase resolution at higher column temperatures. However, increasing column temperature was shown to impart disparaging effects in glycopeptide identification. Herein we further elucidate this trend, describing qualitative and quantitative effects of increased column temperature on glycopeptide identification rates, signal intensity, resolution, and spectral count linear response. Through analysis of enriched bovine and human glycopeptides, species with high mannose and sialylated glycans were shown to most significantly benefit and suffer from high column temperatures, respectively. These results provide insight as to how porous graphitic carbon separations may be appropriately leveraged for glycopeptide identification while raising concerns over quantitative and pseudo-quantitative label free comparisons as temperature changes.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human) Bos Taurus (bovine)

TISSUE(S): Cell Culture

SUBMITTER: Daniel Delafield

LAB HEAD: Lingjun Li

PROVIDER: PXD034354 | Pride | 2023-02-01

REPOSITORIES: Pride

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			Action	DRS
	20210710_GlycEnr_00625_30C_Run1-_1_.mzML	Mzml
	20210710_GlycEnr_00625_30C_Run1.mzid.gz	Mzid
	20210710_GlycEnr_00625_30C_Run1.raw	Raw
	20210710_GlycEnr_00625_30C_Run2-_1_.mzML	Mzml
	20210710_GlycEnr_00625_30C_Run2.mzid.gz	Mzid

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Higher Temperature Porous Graphitic Carbon Separations Differentially Impact Distinct Glycopeptide Classes.

Delafield Daniel G DG Miles Hannah N HN Ricke William A WA Li Lingjun L

Journal of the American Society for Mass Spectrometry 20221130 1

Mass spectrometry-based discovery glycoproteomics is highly dependent on the use of chromatography paradigms amenable to analyte retention and separation. When compared against established stationary phases such as reversed-phase and hydrophilic interaction liquid chromatography, reports utilizing porous graphitic carbon have detailed its numerous advantages. Recent efforts have highlighted the utility in porous graphitic carbon in high-throughput glycoproteomics, principally through enhanced pr ...[more]

PMID: 36450095

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