An integrative strategy for quantitative analysis of the N-glycoproteome in complex biological samples

Wang, Ji; Zhou, Chen; Zhang, Wei; Yao, Jun; Lu, Haojie; Dong, Qiongzhu; Zhou, Haijun; Qin, Lun–Xiu

doi:10.1186/1477-5956-12-4

Cited by 12 publications

(13 citation statements)

References 54 publications

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“…Based on these results, we then sought to identify specific recurrence-related N-linked glycoproteins from the subgroups of ConA-combinable, LCA-combinable or WGA-combinable glycoproteins by using serum samples from a second cohort of 40 patients (set B, including 20 patients with HCC recurrence and 20 patients without HCC recurrence) based on the strategy shown in Supplemental Fig. S3, and previously described by our group 20 . The method is comprised of four steps: digestion of glycoproteins into glycopeptides; lectin affinity chromatography; enzyme-catalyzed 18 O 3 − or 16 O 3 − labeling; and mass spectrum analyses.…”

Section: Resultsmentioning

confidence: 99%

“…The experiments were performed as previously described 20 . Briefly, the serum samples were digested with trypsin at an enzyme-to-substrate ratio of 1:50 (w/w) at 37 °C for 16 h. The digested serum samples were subjected to lectin affinity chromatography for separating the glycopeptides with specific glycan structure using ConA-based, LCH-based, and WGA-based isolation kits.…”

Section: Methodsmentioning

confidence: 99%

“…In a previous study, we described the development of a method for quantifying the N-glycoproteome in blood samples using lectin affinity chromatography combined with enzyme-catalyzed 18 O 3− or 16 O 3− labeling. The potential feasibility of using this approach for the identification of disease-related N-linked glycoproteins using serum samples from HCC patients and healthy individuals was proposed 20 .…”

Section: Introductionmentioning

confidence: 99%

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Core fucosylated glycan-dependent inhibitory effect of QSOX1-S on invasion and metastasis of hepatocellular carcinoma

et al. 2019

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The goal of the present study was to identify glycoproteins associated with the postoperative relapse of hepatocellular carcinoma (HCC) and to investigate their potential role in HCC metastasis. A method for quantitating N-glycoproteome was used to screen for, and identify, recurrence-related N-linked glycoproteins from 100 serum samples taken from patients with early-stage HCC. The prognostic significance of candidate glycoproteins was then validated in 193 HCC tissues using immunohistochemical staining. Serum core fucosylated quiescin sulfhydryl oxidase 1 (cf-QSOX1) was identified as a leading prognostic glycoprotein that significantly correlated with HCC recurrence. Patients with high serum cf-QSOX1 levels had a significantly longer time to recurrence (TTR) as compared with those with low serum cf-QSOX1. As was seen with serum cf-QSOX1, QSOX1 in HCC tissues was further shown to be significantly associated with good patient outcome. Gain-functional and loss-functional analyses of QSOX1-S were performed in vitro and in vivo. QSOX1-S overexpression significantly increased in vitro apoptosis, but decreased the invasive capacity of HCC cells, and reduced lung metastasis in nude mice models bearing human HCC. Furthermore, overexpression of a mutant version of QSOX1-S, which had eliminated the core-fucosylated glycan at Asn-130, showed no demonstrable effect on invasion or metastasis of HCC cells. Our study suggests that serum cf-QSOX1-S and tumor QSOX1 levels are helpful for predicting recurrence in HCC patients, and its core-fucosylated glycan at Asn-130 is critical for the inhibitory effects of QSOX1-S on invasion and metastasis of HCC

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Core fucosylated glycan-dependent inhibitory effect of QSOX1-S on invasion and metastasis of hepatocellular carcinoma

et al. 2019

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“…Glycosylation of proteins changes in a large and dramatic fashion in cancer cells. Glycans become shorter and more negatively charged, and core structures change [24][25][26][27][28]. DSA-FACE is a simple and efficient technology for measuring N-glycan changes in serum.…”

Section: Discussionmentioning

confidence: 99%

Validation of N-glycan markers that improve the performance of CA19-9 in pancreatic cancer

Zhao

Zhou

et al. 2015

Clin Exp Med

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Pancreatic cancer (PC) has a high mortality rate because it is usually diagnosed late. Glycosylation of proteins is known to change in tumor cells during the development of PC. The objectives of this study were to identify and validate the diagnostic value of novel biomarkers based on N-glycomic profiling for PC. In total, 217 individuals including subjects with PC, pancreatitis, and healthy controls were divided randomly into a training group (n = 164) and validation groups (n = 53). Serum N-glycomic profiling was analyzed by DSA-FACE. The diagnostic model was constructed based on N-glycan markers with logistic stepwise regression. The diagnostic performance of the model was assessed further in validation cohort. The level of total core fucose residues was increased significantly in PC. Two diagnostic models designated GlycoPCtest and PCmodel (combining GlycoPCtest and CA19-9) were constructed to differentiate PC from normal. The area under the receiver operating characteristic curve (AUC) of PCmodel was higher than that of CA19-9 (0.925 vs. 0.878). The diagnostic models based on N-glycans are new, valuable, noninvasive alternatives for identifying PC. The diagnostic efficacy is improved by combined GlycoPCtest and CA19-9 for the discrimination of patients with PC from healthy controls.

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“…A disadvantage would be the increased cost of using 18 O-enrich water. Recently, an integrated strategy of lectin affinity separation, followed by passage through immobilized trypsin, quenching of residual trypsin, and PNGase F catalyzed 18 O labeling was introduced [168]. In this case, 18 O labeling of the C-termini (4 Da shift) was done in addition to 18 O labeling of the glycosylation site (to give a total 6 Da shift).…”

Section: Monolithic Reactor Columns In Digestion Of Glycoproteins Andmentioning

confidence: 99%

Development of Monolithic Column Materials for the Separation and Analysis of Glycans

Alla

Stine

2015

Chromatography

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Monolithic column materials offer great advantages as chromatographic media in bioseparations and as solid-supports in biocatalysis. These single-piece porous materials have an interconnected ligament structure that limits the void volume inside the column, thus increasing the efficiency without sacrificing the permeability. The preparation of monolithic materials is easy, reproducible and has available a wide range of chemistries to utilize. Complex, heterogeneous and isobaric glycan structures require preparation methods that may include glycan release, separation and enrichment prior to a comprehensive and site-specific glycosylation analysis. Monolithic column materials aid that demand, as shown by the results reported by the research works presented in this review. These works include selective capture of glycans and glycoproteins via their interactions with lectins, boronic acids, hydrophobic, and hydrophilic/polar functional groups on monolith surfaces. It also includes immobilization of enzymes trypsin and PNGase F on monoliths to digest and deglycosylate glycoproteins and glycopeptides, respectively. The use of monolithic capillary columns for glycan separations through nano-liquid chromatography (nano-LC) and capillary electrochromatography (CEC) and coupling these columns to MS instruments to create multidimensional systems show the potential in the development of miniaturized, high-throughput and automated systems of glycan separation and analysis.

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An integrative strategy for quantitative analysis of the N-glycoproteome in complex biological samples

Cited by 12 publications

References 54 publications

Core fucosylated glycan-dependent inhibitory effect of QSOX1-S on invasion and metastasis of hepatocellular carcinoma

Core fucosylated glycan-dependent inhibitory effect of QSOX1-S on invasion and metastasis of hepatocellular carcinoma

Validation of N-glycan markers that improve the performance of CA19-9 in pancreatic cancer

Development of Monolithic Column Materials for the Separation and Analysis of Glycans

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