JOURNAL OF CHROMATOGRAPHY A
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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
·
分析化学
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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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DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY A
·
2014
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IF 4.258
Qianying Sheng,Xindai Su,Xiuling Li,Yanxiong Ke,Xinmiao Liang
A-saccharide-based stationary phase for hydrophilic interaction liquid chromatography (HILIC) is presented. The method uses carbonyl di-imidazole (CDI) as a cross-linker for dextran in aqueous solution. Different from the traditional immobilized saccharide stationary phase, this method was using a simple way to bond the high molecular of polysaccharide immobilized on the silica gel. The new method avoided the time-consuming process but had a very considerable loading result. With silica-based surface bonded, it will be possible to develop well-defined surface modifications that promote the hydrophilic with dextran and high mechanical strength with silica. For all tested compounds, including polar compounds and carbohydrates, this dextran-bonded stationary phase performed well in terms of separation efficiency and column stability, and the retention mechanism matched that of typical HILIC retention. Moreover, good selectivity was achieved in the separation of oligosaccharides and glycopeptides.
研究领域:
分析化学
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色谱学
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DOI
PubMed
高影响因子
MICROCHIMICA ACTA
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2021
·Vol. 188
·期 10
·1-10
·
IF 5.833
Wang Baichun,Liu Jianmu,Yan Yinghua,Ding Chuan-Fan,Tang Keqi
A novel type of boric acid–functionalized magnetic covalent organic framework (mCOF) with polyethyleneimine (PEI) as a linker (denoted as mCOF@PEI@B(OH) 2 ) has been prepared through a post-synthesis strategy, which points out an achievable path for the construction of boronic acid–functionalized COFs. Based on the boric acid chemistry, the obtained core-shell structured mCOF@PEI@B(OH) 2 can selectively isolate glycopeptides through the modified boronic acid groups. The mCOF@PEI@B(OH) 2 exhibits excellent performance with good reusability (ten cycles), low detection limit (0.5 fmol·μL −1 ), size-exclusion effect, and relatively high loading capacity (80 μg·mg −1 ), recovery yield (94.9 ± 2.8%), and selectivity (HRP digests:BSA digests = 1:500). Detection is done by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). In addition, 37 endogenous glycopeptides are captured from human saliva with mCOF@PEI@B(OH) 2 , providing effective proofs for its capability to capture low-abundance glycopeptides from actual biological samples. Graphical abstract
研究领域:
材料科学
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分析化学
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生物化学
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质谱分析
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DOI
PubMed
高影响因子
Science China-Chemistry
·
2015
·Vol. 58
·期 6
·1056-1064
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IF 1.695
Zheng Jiangnan,Lin Zian,Zhang Lan,Yang Huanghao
In this work, we demonstrate for the first time, a method to synthesize phenylboronic acid-Fe 3 O 4 @polydopamine (Fe 3 O 4 @PDA-PBA) magnetic microspheres via the combination of mussel-inspired polydopamine coating and click chemistry. Uniform-size and core-shell structured Fe 3 O 4 @PDA-PBA magnetic microspheres with a core diameter of ∼240 nm and a shell thickness of ∼13 nm were obtained as identified by the characterization of the morphology, structure and composition of the synthesized microspheres. We evaluated the selectivity and binding capacity of the Fe 3 O 4 @PDA-PBA magnetic microspheres by using standard glycoproteins (ovalbumin, immunoglobulin G and catalase) and nonglycoproteins (human serum albumin, bovine hemoglobin, myoglobin, lysozyme, and ribonuclease A) as model proteins. Adsorption experiments, SDS-PAGE and mass spectrometry analysis demonstrated that the Fe 3 O 4 @PDA-PBA magnetic microspheres had much high binding capacity and selectivity for glycoproteins/glycopeptides compared to nonglycoproteins/nonglycopeptides. In addition, the practicability of the Fe 3 O 4 @PDA-PBA magnetic microspheres was further assessed by selective capture of glycoproteins from healthy human serum. The good results demonstrated its potential in glycoproteome analysis.
研究领域:
生物化学
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分析化学
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材料科学
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DOI
高影响因子
TALANTA
·
2021
·
IF 6.057
Zehu Xie,Yinghua Yan,Keqi Tang,Chuan-Fan Ding
In this study, a novel type of covalent organic framework (COF) material rich in boronic acid sites was prepared through post-synthesis modification (TbBD@PEI@Au@4-MPBA). The surface of COF material had abundant carboxylic acid groups, which could bind a large amount of polyethyleneimine (PEI) through electrostatic interaction . At the same time, the amino groups on the PEI can be grafted with Au nanoparticles (Au NPs) in situ, and then 4-mercaptophenylboronic acid (4-MPBA) was modified by the reaction of Au and sulfhydryl groups. The massive grafting of boronic acid groups made the material's enrichment effect on glycopeptides expected. The results of experiments indicated that the composite material has high sensitivity (5 amol μL −1 ) and selectivity (1:1000). In addition, the material has outstanding stability and reusability, with a load capacity of about 100 mg g −1 and a recovery of 99.3 ± 2.2%. What's more, after enriched by TbBD@PEI@Au@4-MPBA, 56 endogenous glycopeptides from fresh human saliva were detected by MALDI-TOF MS, 56 unique glycopeptides corresponding to 31 glycoproteins from human saliva and 513 unique glycopeptides corresponding to 208 glycoproteins from serum of throat cancer patient were detected by nano-LC-MS/MS, respectively, which was expected to be applied to glycoproteomics research.
研究领域:
生物化学
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医学诊断
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DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2024
·
IF 2.8
Luyao Yu,Xiaoqian Chen,Yingxia Guo,Jiansong You,Meiyun Shi,Yalin Xi,Lei Yin
An ultra-high performance liquid chromatography tandem mass spectrometry cubed (UHPLC/MS 3 ) assay coupled with protein precipitation and counter-extraction for detection of tricyclic glycopeptide vancomycin in human plasma was established and validated in this study. After protein precipitation and counter-extraction with dichloromethane, chromatographic separation of vancomycin and norvancomycin were performed on a reversed phase column (XBridge Peptide BEH C 18 column, 2.1 × 100 mm I.D, 3.5 μm). The transition (parent ions → fragment ions → further fragment ions) at m / z 725.3 → 144.1 → 100.1 was used for quantification of vancomycin. The transition (parent ions → fragment ions) at m / z 718.3 → 144.2 was used for detection of norvancomycin. The linear range of the developed analytical method for quantification of vancomycin was 0.5–100 µg/mL (r = 0.9989). The range of intra- and inter-day precisions of the assay among low, medium and high concentrations is between 1.88 % and 6.33 %. The sensitivity of the analytical method was significantly improved by using MS 3 technique as monitoring mode and counter-extraction with dichloromethane followed by protein precipitation as sample processing assay. The developed UHPLC/MS 3 assay was successfully applied for clinical therapeutic drug monitoring (TDM) of vancomycin in 45 human plasma samples.
研究领域:
分析化学
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临床药理学
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药物代谢动力学
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DOI
PubMed
高影响因子
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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产品关联
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
ACS Applied Polymer Materials
·
2025
·
IF 4.7
Penghui Wang,Yingying Yang,Yudong Yin,Hua Zhang,Tianqi Shi,Wenjie Zhang,Huiyun Liao,Shuang Li,Xiaoyan Tan,Zheng Yao,Bo Chi
Hydrogel microspheres have attracted extensive attention as the key carriers for drug delivery and cell culture. The traditional microspheres are limited in their application in the regenerative medicine fields due to their closed surface structure and biological inertness. To this end, this study used polylactic acid and selenocysteine modified hyaluronic acid through microfluidic technology and a microsphere surface modification strategy to prepare a hydrogel porous microsphere with reactive oxygen species (ROS) responsive properties and achieved effective removal of ROS through Se–Se rupture. It was then incorporated into a glycopeptide hydrogel, cross-linked by polyglutamic acid and hyaluronic acid, incorporating poloxamer. The glycopeptide hydrogels/microspheres with temperature response characteristics were prepared to reconstruct the dynamic microenvironment required for the repair and regeneration of damaged tissues. The interaction between dynamic and static bonds in the hydrogel system gives the microsphere composite hydrogel excellent injectability, mechanical strength, structural stability, organizational adaptability, and biocompatibility. It can be used as a tissue implant material in the field of tissue regeneration and cell microenvironment reconstruction with a broad application potential.
研究领域:
生物材料学
·
再生医学
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产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）