JOURNAL OF CHROMATOGRAPHY A
·
2016
·
IF 3.926
Yiman Zhao,Yajing Chen,Zhichao Xiong,Xudong Sun,Quanqing Zhang,Yangyang Gan,Lingyi Zhang,Weibing Zhang
In consideration of the close connection between glycopeptides and human diseases, the efficient method to separate and enrich glycopeptides from complex biological samples is urgently required. In the work, we developed a magnetic zwitterionic-hydrophilic material for highly effective separation and analysis of glycopeptides from complex samples. The Fe 3 O 4 particles were covered with a thick layer of polymer by one-step reflux-precipitation polymerization (RPP), subsequently decorated by Au nanoparticles (Au NPs) through in situ reduction and finally modified with zwitterionic groups. The abundant zwitterionic sites facilitate the selective enrichment of glycopeptides. Besides, the prepared Fe 3 O 4 @PGMA@Au- l -cys showed high detection sensitivity (5 fmol IgG digest), approving enrichment capacity (75 mg g −1 ), satisfactory enrichment recovery (89.8%), and great performance in the analysis and profiling of low-abundance N-linked glycopeptides. Furthermore, the prepared material was employed in the enrichment of glycopeptides in intricate biological samples, and 774 unique N -glycosylation sites from 411 N -glycosylated proteins were reliably identified in three replicate analyses of a 75 μg protein sample extracted from mouse liver, suggesting wide application prospect in glycoproteomics.
研究领域:
分析化学
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蛋白质组学
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医学
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DOI
PubMed
高影响因子
ANALYST
·
2017
·Vol. 142
·期 17
·3212-3218
·
IF 3.885
Yu-Fang Ma,Fang Yuan,Xiao-Hui Zhang,Ying-Lin Zhou,Xin-Xiang Zhang
The enrichment of glycopeptides plays an important role in glycoproteomics. In this paper, a covalent-organic framework called TpPa-1, synthesized by the Schiff base reaction of 1,3,5-triformylphloroglucinol and paraphenylenediamine, was first successfully utilized as a hydrophilic porous material for N-linked glycopeptide enrichment. Using this material, interference from non-glycopeptides could be efficiently eliminated, which facilitated the mass spectrometry detection of glycopeptides. By capturing N-linked glycopeptides from tryptic digests of human IgG, our method was proved to have high sensitivity at the femtomole level. And it showed superior selectivity for glycopeptides even when non-glycopeptides were 1000 times more concentrated. Due to the strong covalent bonds, this material possessed good stability and could be repeatedly used for at least 10 times. The ultra-low mass density and abundant binding sites also provided it with high binding capacity (178 mg g−1, IgG/TpPa-1). Moreover, N-linked glycopeptides were easily enriched by this material from only 10 μL human serum, which demonstrated its potential in pretreatment of complex biological samples.
研究领域:
蛋白质组学
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分析化学
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生物医学
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产品关联
DOI
PubMed
高影响因子
ANALYST
·
2022
·Vol. 147
·期 11
·2425-2432
·
IF 5.227
Haozhou Jin,Wenqing Gao,Rong Liu,Jiaqian Yang,Shun Zhang,Renlu Han,Jing Lin,Sijia Zhang,Jiancheng Yu,Keqi Tang
Protein glycosylation is of great significance in various physiological processes. Nevertheless, it remains a huge challenge to identify glycopeptides in complex biosamples by the direct mass spectrometry analysis due to the low ion efficiency and low abundance of glycopeptides. In this study, a novel hydrogel (denoted as ZIF-8/SAP) with a stable three-dimensional (3D) network structure and excellent hydrophilicity was successfully fabricated to capture glycopeptides with high efficiency. Owing to the unique characteristics, ZIF-8/SAP exhibited great selectivity (1 : 1000), great sensitivity (1 fmol μL−1), large binding capacity (300 mg g−1) and satisfactory reusability (seven cycles). Inspired by the great enrichment performance of the as-prepared material toward glycopeptides, ZIF-8/SAP was further applied to capture glycopeptides from a real human serum sample. The experimental results demonstrated that 217 N-glycosylation sites were identified in 283 N-glycopeptides, corresponding to 95 glycoproteins identified from 10 μL human serum by the nano-LC-MS/MS analysis, revealing the great potential of the novel ZIF-8/SAP hydrogel for glycopeptide enrichment and glycoproteomic research.
研究领域:
蛋白质组学
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生物化学
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产品关联
DOI
PubMed
高影响因子
Journal of Functional Biomaterials
·
2024
·Vol. 15
·期 7
·185
·
IF 5
Yu Wu,Sen Xu,Fengjuan Ding,Weibing Zhang,Haiyan Liu
In this work, a new type of FE-1 COF material is prepared by a reversible imine condensation reaction with diaminoferrocene and diaminodiformaldehyde as materials. The material is connected by imine bonds to form a COF skeleton, and the presence of plenty of nitrogen-containing groups gives the material good hydrophilicity; the presence of metal Fe ions provides the material application potential in the enrichment of phosphopeptides. According to the different binding abilities of N-glycopeptide and phosphopeptide on FE-1 COF, it can simultaneously enrich N-glycopeptide and phosphopeptide through different elution conditions to realize its controllable and selective enrichment. Using the above characteristics, 18 phosphopeptides were detected from α-casein hydrolysate, 8 phosphopeptides were detected from β-casein hydrolysate and 21 glycopeptides were detected from IgG hydrolysate. Finally, the gradual elution strategy was used; 16 phosphopeptides and 19 glycopeptides were detected from the α-casein hydrolysate and IgG hydrolysate. The corresponding glycopeptides and phosphopeptides were identified from the human serum. It proves that the FE-1 COF material has a good enrichment effect on phosphopeptides and glycopeptides.
研究领域:
分析化学
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蛋白质组学
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生物化学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY A
·
2019
·
IF 3.858
Yong Zhang,Hongyu Jing,Bo Meng,Xiaohong Qian,Wantao Ying
The facile enrichment of glycopeptides or glycoproteins poses great challenges for glycoproteomic research. In this study, a novel hydrophilic material, named zwitterionic hydrophilic L-cysteine derivatized straticulate-C 3 N 4 composites (LCAC), were synthesized and evaluated for the enrichment of N-glycopeptides. LCAC exhibited good biocompatibility, excellent hydrophilicity and selectivity, by virtue of the large surface of C 3 N 4 and the zwitterionic property offered by cysteine. LCAC demonstrated excellent performance for N-glycopeptide enrichment with the sensitivity of 0.033 fmol/µL, selectivity of 1:100, and high recovery rate (∼85%). The performance of LCAC was demonstrated by the identification of 35 N-glycopeptides from IgG, as well as capturing 1809 human urine N-glycopeptides corresponding to 876 N-glycoproteins. Comparing the LCAC with our developed phenylboronic acid functionalized material showed a certain complementary due to the different binding mechanism. The simple production and enhanced hydrophilic properties make the material a promising choice for glycoproteomics researches.
研究领域:
蛋白质组学
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分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2019
·
IF 6.35
Luwei Zhang,Shujuan Ma,Yao Chen,Yan Wang,Junjie Ou,Hiroshi Uyama,Mingliang Ye
In the study of glycoproteomics with mass spectrometry, certain pretreatments of samples are required for eliminating the interference of nonglycopeptides and improving the efficiency of glycopeptides detection. Although hydrophilic interaction chromatography (HILIC) has been developed for enrichment of glycosylated peptides, a plethora of hydrophilic materials always suffered from large steric hindrance, great cost, and difficulty with modifications of high-density hydrophilic groups. In this work, a 1 mm thick biomimetic honeycomb chitosan membrane (BHCM) with honeycomb-like accessible macropores was directly prepared by the freeze-casting method as an adsorbent for HILIC. The N-glycopeptides from trypsin digests of immunoglobulin G (IgG), mixture of IgG and bovine serum albumin (BSA), and serum proteins were enriched using this material and compared with a commercial material ZIC-HILIC. The biomimetic membrane could identify as many as 32 N-glycopeptides from the IgG digest, exhibiting high sensitivity (about 50 fmol) and a wide scope for glycopeptide enrichment. A molar ratio of IgG trypsin digest to bovine serum albumin trypsin digest as low as 1/500 verified the outstanding specificity and efficiency for glycopeptide enrichment. In addition, 270 unique N-glycosylation sites of 400 unique glycopeptides from 146 glycosylated proteins were identified from the triplicate analysis of 2 μL human serum. Furthermore, 48 unique O-glycosylation sites of 278 unique O-glycopeptides were identified from the triplicate analysis of 30 μg deglycosylated fetuin digest. These results indicated that the chitosan-based membrane prepared in this work had great potential for pretreatment of samples in glycoproteomics.
研究领域:
生物化学
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蛋白质组学
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分析化学
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2021
·Vol. 188
·期 8
·1-9
·
IF 5.833
Gao Wenjie,Bai Yu,Liu Huwei
Protein glycosylation plays pivotal role in a variety of biological processes and has association with many diseases. The highly efficient glycopeptide enrichment is essential for the mass spectrometry–based glycoproteome research to reduce interference from non-glycopeptides. In this study, novel glutathione-functionalized two-dimensional cobalt sulfide nanosheets (Co-S@Au-GSH) were synthesized for rapid and highly effective enrichment of glycopeptides. By using this nanomaterial, 34 and 21 N-glycopeptides were effectively captured from human serum immunoglobulin G (IgG) and horseradish peroxidase (HRP) digests, respectively. In addition, the Co-S@Au-GSH showed remarkable performance in N-glycopeptide extraction with high selectivity (HRP: BSA = 1:500), low limit of detection (0.5 fmol/μL), high binding capacity (150 mg/g), good reusability, and great robustness. Moreover, it was successfully applied in complex serum samples, demonstrating its excellent enrichment performance. These results indicated that this nanomaterial has great potential in complicated practice samples in glycoproteome determination. Graphical abstract
研究领域:
蛋白质组学
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生物化学
·
分析化学
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2020
·
IF 5.977
Ruizhi Tang,Yang Yu,Jing Dong,Yating Yao,Shujuan Ma,Junjie Ou,Mingliang Ye
In bottom-up strategy, specific enrichment of glycopeptides and phosphopeptides from complicated biological samples is a prerequisite for efficient identifying glycosylation and phosphorylation by mass spectrometry . Although there were a plethora of materials used as either hydrophilic interaction liquid chromatography (HILIC) or immobilized metal affinity chromatography (IMAC) adsorbents, even several bifunctional materials for simultaneous enrichment of glycopeptides and phosphopeptides, most of them are not easily commercialized as many other well-performing adsorbents due to the complicated preparation process. In our case, a one-step modification strategy was developed to prepare bifunctional adsorbents for HILIC and IMAC, employing O -phospho- l -serine as the modifier and poly(GMA- co -EDMA) microspheres, a kind of macroporous adsorption resin (MAR) with epoxy groups , as the matrix. The MARs were directly modified with O -phospho- l -serine under facile condition for HILIC strategy and further chelated with Ti 4+ for IMAC strategy. A total of 522 unique N -glycopeptides and 442 unique N -glycosylation sites mapped to 275 N -glycoproteins was identified from HeLa cell proteins, showing excellent enrichment efficiency in HILIC. Additionally, 3141 unique phosphopeptides were unambiguously identified from 200 μg of digest of HeLa cell proteins, demonstrating great enrichment efficiency in IMAC. Moreover, these materials have been successfully applied in the analysis of multiple biological samples including human serum and milk, demonstrating their feasibility for real sample applications and potential business value.
研究领域:
蛋白质组学
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生物化学
·
分析化学
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DOI
高影响因子
JOURNAL OF CHROMATOGRAPHY A
·
2024
·
IF 3.8
Liang Lai,Mengyao Zhang,Yuexiang Chang,Minyi Li,Jacques Crommen,Jia-Huan Qu,Dongsheng Xu,Zhengjin Jiang
The structure of zwitterion has great impact on the separation properties of zwitterionic hydrophilic stationary phases. To better understand the role of anionic groups of zwitterions, a novel carboxybetaine-based zwitterionic monolithic column was first prepared through thermo-initiated copolymerization of functional monomer (3-acrylamidopropyl)-dimethyl-(2-carboxymethyl) ammonium (CBAA) and crosslinker ethylene dimethacrylate (EDMA) within 100 μm ID capillary. The optimal poly(CBAA- co -EDMA) monolithic column exhibited satisfactory mechanical and chemical stability, good repeatability, high column efficiency (96,000 plates / m), and excellent separation performance for different classes of polar compounds (i.e., phenols, monophosphate nucleotides, urea and allantoin). A comparative study was then performed among three zwitterionic hydrophilic stationary phases containing different anionic groups, i.e. poly(CBAA- co -EDMA) (carboxybetaine), poly(2-{2-(methacryloyloxy) ethyldimethylammonium}ethyl n-butyl phosphate- co -EDMA) (phosphocholine), and poly(N,N-dimethyl-N-(3-methacrylamidopropyl)-N-(3-sulfopropyl) ammonium betaine- co -EDMA) (sulfobetaine) using benzoic acid derivatives, amine compounds, nucleobases and nucleosides as model analytes. The carboxybetaine-based monolithic column exhibited much higher positive zeta-potential and hydrophilicity, which endows it with a stronger retention capacity for acidic and neutral compounds, but sulfobetaine-based monolithic column exhibited much higher selectivity and retention capacity for the amines. Moreover, their enrichment efficiencies for N- glycopeptides were also evaluated based on their different hydrophilicity, and it was observed that the poly(CBAA- co -EDMA) monolithic material captured 4-8 times more N- glycopeptides compared to the other two materials.
研究领域:
分析化学
·
色谱技术
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产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2019
·
IF 5.256
Qiqin Wang,Qiaoxuan Zhang,Hao Huang,Pengwei Zhao,Lingjue Sun,Kun Peng,Xiao Liu,Meng Ruan,Huikai Shao,Jacques Crommen,Pei Yu,Zhengjin Jiang
In this study, a series of zwitterionic phosphorylcholine functionalized monolithic columns were fabricated via the thermally initiated co-polymerization of 2-methacryloyloxyethyl phosphorylcholine (MPC) and different hydrophilic crosslinkers, including 1,4-bis(acryloyl)piperazine (PDA), N,N′-methylenebisacrylamide (MBA) and 2-(methacryloyloxy)ethyl-[N-(2-methacryloyloxy)ethyl]phosphorylcholine (MMPC). The physicochemical and chromatographic properties of these MPC functionalized monoliths, including column morphology, pore size distribution , permeability, column efficiency, retention mechanism and ζ-potential analysis, were systematically compared. Furthermore, the influence of the crosslinker on the chromatographic performance of these MPC functionalized monoliths was evaluated. The chromatographic results indicate that the polarity of MPC functionalized monoliths may be related to the polarity of the crosslinker, which further affects the column selectivity and efficiency. A particularly high column efficiency (88,000 plates/m) was obtained on the novel poly(MPC- co -MMPC) monolith at optimum linear velocity using thiourea as test analyte. Compared to the poly(MPC- co -MBA) and poly(MPC- co -PDA) monoliths, the poly(MPC- co -MMPC) monolith exhibited higher separation selectivity for polar analytes, including nucleobases , nucleosides and benzoic acid derivatives. Moreover, 24 N -glycopeptides could be detected after enrichment with the poly(MPC- co -MMPC) versus 19 and 10 N -glycopeptides with the poly(MPC- co -MBA) and poly(MPC- co -PDA) monoliths, and no N -glycopeptide without enrichment. Therefore, MMPC has a great potential as a new and alternative hydrophilic crosslinker for the development of zwitterionic polymeric monoliths.
研究领域:
分析化学
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生物化学
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产品关联
DOI
PubMed
高影响因子
Nanoscale
·
2014
·Vol. 7
·期 7
·3100-3108
·
IF 6.739
Yajing Chen,Zhichao Xiong,Lingyi Zhang,Jiaying Zhao,Quanqing Zhang,Li Peng,Weibing Zhang,Mingliang Ye,Hanfa Zou
Highly selective and efficient capture of glycosylated proteins and peptides from complex biological samples is of profound significance for the discovery of disease biomarkers in biological systems. Recently, hydrophilic interaction liquid chromatography (HILIC)-based functional materials have been extensively utilized for glycopeptide enrichment. However, the low amount of immobilized hydrophilic groups on the affinity material has limited its specificity, detection sensitivity and binding capacity in the capture of glycopeptides. Herein, a novel affinity material was synthesized to improve the binding capacity and detection sensitivity for glycopeptides by coating a poly(2-(methacryloyloxy)ethyl)-dimethyl-(3-sulfopropyl) ammonium hydroxide (PMSA) shell onto Fe3O4@SiO2 nanoparticles, taking advantage of reflux-precipitation polymerization for the first time (denoted as Fe3O4@SiO2@PMSA). The thick polymer shell endows the nanoparticles with excellent hydrophilic property and several functional groups on the polymer chains. The resulting Fe3O4@SiO2@PMSA demonstrated an outstanding ability for glycopeptide enrichment with high selectivity, extremely high detection sensitivity (0.1 fmol), large binding capacity (100 mg g−1), high enrichment recovery (above 73.6%) and rapid magnetic separation. Furthermore, in the analysis of real complicated biological samples, 905 unique N-glycosylation sites from 458 N-glycosylated proteins were reliably identified in three replicate analyses of a 65 μg protein sample extracted from mouse liver, showing the great potential of Fe3O4@SiO2@PMSA in the detection and identification of low-abundance N-linked glycopeptides in biological samples.
研究领域:
生物化学
·
材料科学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2017
·
IF 4.95
Qiqin Wang,Huihui Wu,Kun Peng,Hanying Jin,Huikai Shao,Yuqiang Wang,Jacques Crommen,Zhengjin Jiang
In this research, a hydrophilic polymeric monolith containing choline phosphate (CP) was fabricated through the thermally initiated free-radical polymerization of 2-{2-(methacryloyloxy) ethyldimethylammonium}ethyl n -butyl phosphate (MBP) with ethylene dimethacrylate (EDMA) using isopropanol and tetrahydrofuran as the porogenic system. Excellent mechanical strength, permeability, stability and reproducibility were obtained on the optimized poly(MBP- co -EDMA) monolith. Column efficiency as high as 107,500 plates/m was achieved for the analysis of thiourea at a flow velocity of 0.2 mm/s on the monolith. Both hydrophilic and electrostatic interactions were observed for the retention of charged analytes on the poly(MBP- co -EDMA) monolith. It is worth noting that the resulting monolith exhibits higher selectivity and efficiency than the classical 2-(methacryloyloxy)ethyl phosphorylcholine functionalized polymeric monolith for the chromatographic separation of polar analytes. Also, the novel monolithic column was successfully applied to enrich N -glycopeptides from tryptic digest of human IgG. In a word, the versatile MBP functionalized polymeric monolith not only opens up interesting possibilities for reverse zwitterionic CP derivative-based polymeric materials in separation science , but also represents a promising hydrophilic interaction chromatography stationary phase.
研究领域:
分析化学
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生物化学
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产品关联
DOI
PubMed
高影响因子
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