GlyQ-IQ: Glycomics Quintavariate-Informed Quantification with High-Performance Computing and GlycoGrid 4D Visualization

Glycomics quintavariate-informed
quantification (GlyQ-IQ) is a
biologically guided glycomics analysis tool for identifying N-glycans
in liquid chromatography–mass spectrometry (LC–MS) data.
Glycomics LC–MS data sets have convoluted extracted ion chromatograms
that are challenging to deconvolve with existing software tools. LC
deconvolution into constituent pieces is critical in glycomics data
sets because chromatographic peaks correspond to different intact
glycan structural isomers. The biological targeted analysis approach
offers several key advantages to traditional LC–MS data processing. A priori glycan information about the individual target’s
elemental composition allows for improved sensitivity by utilizing
the exact isotope profile information to focus chromatogram generation
and LC peak fitting on the isotopic species having the highest intensity.
Glycan target annotation utilizes glycan family relationships and
in source fragmentation in addition to high specificity feature LC–MS
detection to improve the specificity of the analysis. The GlyQ-IQ
software was developed in this work and evaluated in the context of
profiling the N-glycan compositions from human serum LC–MS
data sets. A case study is presented to demonstrate how GlyQ-IQ identifies
and removes confounding chromatographic peaks from high mannose glycan
isomers from human blood serum. In addition, GlyQ-IQ was used to generate
a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid
chromatography–tandem mass spectrometry (nESI-LC–MS/MS)
data set. A total of 156 glycan compositions and 640 glycan isomers
were detected from a single sample. Over 99% of the GlyQ-IQ glycan-feature
assignments passed manual validation and are backed with high-resolution
mass spectra.