N‐Glycan Analysis by Ultra‐Performance Liquid Chromatography and Capillary Gel Electrophoresis with Fluorescent Labeling

Lauc, Gordan; Trbojević‐Akmačić, Irena

doi:10.1002/cpps.95

Cited by 22 publications

(18 citation statements)

References 8 publications

(6 reference statements)

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“…The basic characteristics of the cohorts are given in Table 1. IgG glycosylation was analysed using a well-established reliable liquid chromatography method for IgG N-glycan analysis with con rmed reproducibility and robustness 30,31 , and which has already been successfully employed to detect signi cant changes in glycosylation in many different diseases (such as COVID-19 32 , thyroid disease 33 , multiple sclerosis 34 , rheumatoid arthritis 35 , cardiovascular disease 36 and systemic lupus erythematosus 37 ). IgG glycans were separated into 24 chromatographic peaks (GP1-GP24) and glycans corresponding to each individual peak are shown in Figure 1.…”

Section: Resultsmentioning

confidence: 99%

Accelerated biological aging in people with Down syndrome with full and segmental trisomy 21 begins in childhood as revealed by immunoglobulin G glycosylation

Cindrić

Vučković

Koschut

et al. 2021

Preprint

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Cells from people with Down syndrome (DS) show faster accumulation of DNA damage and epigenetic aging marks. Causative mechanisms remain un-proven and hypotheses range from amplified chromosomal instability to actions of several supernumerary chromosome 21 genes. Plasma immunoglobulin G (IgG) glycosylation profiles are established as a reliable predictor of biological and chronological aging. We performed IgG glycan profiling of n=246 individuals with DS (208 adults and 38 children) from three European populations and compared these to age-, sex- and demography-matched general populations. We uncovered very significantly increased IgG glycosylation aging marks associated with DS. Average levels of IgG glycans without galactose (G0) and those with two galactoses (G2) as a function of age in persons with DS corresponded to levels detected in 19 years older euploid individuals. Some aging marks were significant already in children with DS. Remarkably, the IgG glycan profiles of a child with segmental duplication of only 31 genes on chromosome 21 had values similar to those of age-matched DS children, outside the normal children’s range. This is the first non-epigenetic evidence of accelerated systemic biological aging in DS, suggesting it begins very early in childhood. It points to a causative contribution of the overdose of genes in a short segment of chromosome 21, not previously linked to accelerated aging, opening a route to discovery of hitherto unrecognised mechanisms.

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Section: Resultsmentioning

confidence: 99%

Accelerated biological aging in people with Down syndrome with full and segmental trisomy 21 begins in childhood as revealed by immunoglobulin G glycosylation

Cindrić

Vučković

Koschut

et al. 2021

Preprint

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“…IgG N‐glycans were analyzed in accordance with the previously published high‐throughput protocol [23] with slight modifications: Before denaturation, total volume (1170 μL) of neutralized IgG eluates (mean IgG concentration 0.01 μg/μL) was dried down in a vacuum concentrator and desalted: 800 μL of cold (–20°C) methanol (J. T. Baker, USA) was added to each sample, the samples were resuspended by pipetting and centrifuged at 2000 × g for 15 min. 780 μL of supernatant was removed, the procedure was repeated and the samples were finally dried down in a vacuum concentrator.…”

Section: Figurementioning

confidence: 99%

N‐glycosylation analysis of mouse immunoglobulin G isolated from dried blood spots

Patenaude

Erhardt

Hennig³

et al. 2020

Electrophoresis

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The association of immunoglobulin G (IgG) glycosylation changes with various human diseases and physiological conditions is well established. Since the mechanistical explanation of the regulation of IgG glycosylation and its functional role in these various states is still missing, the eyes of the biomedical community are now turned towards animal models, which enable intervention studies necessary for conclusions on causality. Mice are recognized and used as a good experimental model for human IgG glycosylation. However, smaller blood volumes, low IgG concentrations at young ages (which are most often used in mice experiments) and multiple sampling protocols during the course of longitudinal studies would profit from a robust workflow for mouse IgG glycome analysis from minute amounts of starting material, collected through a simple sampling procedure. For this purpose, we have developed a protocol for analysis of total N‐glycans of IgG isolated from mouse dried blood spots (DBS), which we report here. We show that mouse DBS are a good source of material for IgG N‐glycan analysis by multiplexed capillary gel electrophoresis with laser‐induced fluorescence (xCGE‐LIF).

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“…only IgG galactosylation was estimated from the total plasma glycome profile, prevented us from the detailed characterization of the estrogen effect on IgG glycosylation. In the present study, we aimed at a better understanding of the estrogen role in the regulation of IgG glycosylation, therefore we reanalyzed samples from the previous intervention study ( 14 ) using state-of-the-art glycoprofiling technologies ( 15 ). We first defined the components of IgG glycome affected by estradiol (E 2 ).…”

Section: Introductionmentioning

confidence: 99%

Effects of Estradiol on Immunoglobulin G Glycosylation: Mapping of the Downstream Signaling Mechanism

et al. 2021

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Glycans attached to immunoglobulin G (IgG) directly affect this antibody effector functions and regulate inflammation at several levels. The composition of IgG glycome changes significantly with age. In women, the most notable change coincides with the perimenopausal period. Aiming to investigate the effect of estrogen on IgG glycosylation, we analysed IgG and total serum glycomes in 36 healthy premenopausal women enrolled in a randomized controlled trial of the gonadotropin-releasing hormone analogue (GnRHAG) leuprolide acetate to lower gonadal steroids to postmenopausal levels and then randomized to transdermal placebo or estradiol (E2) patch. The suppression of gonadal hormones induced significant changes in the IgG glycome, while E2 supplementation was sufficient to prevent changes. The observed glycan changes suggest that depletion of E2 primarily affects B cell glycosylation, while liver glycosylation stays mostly unchanged. To determine whether previously identified IgG GWAS hits RUNX1, RUNX3, SPINK4, and ELL2 are involved in downstream signaling mechanisms, linking E2 with IgG glycosylation, we used the FreeStyle 293-F transient system expressing IgG antibodies with stably integrated CRISPR/dCas9 expression cassettes for gene up- and downregulation. RUNX3 and SPINK4 upregulation using dCas9-VPR resulted in a decreased IgG galactosylation and, in the case of RUNX3, a concomitant increase in IgG agalactosylation.

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N‐Glycan Analysis by Ultra‐Performance Liquid Chromatography and Capillary Gel Electrophoresis with Fluorescent Labeling

Cited by 22 publications

References 8 publications

Accelerated biological aging in people with Down syndrome with full and segmental trisomy 21 begins in childhood as revealed by immunoglobulin G glycosylation

Accelerated biological aging in people with Down syndrome with full and segmental trisomy 21 begins in childhood as revealed by immunoglobulin G glycosylation

N‐glycosylation analysis of mouse immunoglobulin G isolated from dried blood spots

Effects of Estradiol on Immunoglobulin G Glycosylation: Mapping of the Downstream Signaling Mechanism

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