ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2022
·Vol. 414
·期 5
·1881-1890
·
IF 4.478
Wu Yike,Liu Yuanyuan,Shang Zhengjun,Liu Xin,Xu Yong,Liu Wenlan
Aberrant protein N-glycosylation is one of the hallmarks of malignancy, while its role in oral cancer (OC) progression is still poorly understood making it a perfect biomarker for early-stage cancer. In this work, we describe a MALDI-MS-based N-glycomic analysis with multivariate data analysis approach for exploration and classification of the patients with OC. Various statistical analyses were used to determine differential N-glycome profiles. There were 8 glycan structures significantly increased ( p < 0.05) in all OC-derived microvesicles (MVs). Importantly, the increases in these glycans were significantly greater in the early stage of OC than those in the advanced stage of OC. Additionally, N-glycomic profiles from the serum were also performed, demonstrating a significant change of 11 glycan structures ( p < 0.05). Our data highlighted the effectiveness approach of MALDI-MS-based MV N-glycomic profiling on OC screening, and presented strong potential for further identifying targeted glycoproteomics signatures of the cancer.
研究领域:
生物化学
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肿瘤学
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分子生物学
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DOI
PubMed
高影响因子
Scientific Reports
·
2017
·Vol. 7
·期 1
·1-12
·
IF 4.259
Wu Zhaoguan,Li Henghui,Zhang Qiwei,Liu Xin,Zheng Qi,Li Jianjun
O -Acetylation of sialic acid in protein N-glycans is an important modification and can occur at either 4-, 7-, 8- or 9-position in various combinations. This modification is usually labile under alkaline reaction conditions. Consequently, a permethylation-based analytical method, which has been widely used in glycomics studies, is not suitable for profiling O -acetylation of sialic acids due to the harsh reaction conditions. Alternatively, methylamidation can be used for N-glycan analysis without affecting the base-labile modification of sialic acid. In this report, we applied both permethylation and methylamidation approaches to the analysis of O -acetylation in sialic acids. It has been demonstrated that methylamidation not only stabilizes sialic acids during MALDI processing but also allow for characterization of their O -acetylation pattern. In addition, LC-MS/MS experiments were carried out to distinguish between the O -acetylated glycans with potential isomeric structures. The repeatability of methylamidation was examined to evaluate the applicability of the approach to profiling of O -acetylation in sialic acids. In conclusion, the combination of methylamidation and permethylation methodology is a powerful MALDI-TOF MS-based tool for profiling O -acetylation in sialic acids applicable to screening of N-glycans.
研究领域:
生物化学
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分析化学
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糖组学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2019
·
IF 6.393
Shuzhen Dou,Ning Li,Xueyun Wang,Qunyan Zhu,Zhongshun Wang,Nan Lu
Rapid quantification of peptides by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is a challenge. Herein, we prepared a super-hydrophobic poly(methyl methacrylate) (PMMA) structured chip for quantifying peptides without internal standard by improving the detection reproducibility and sensitivity. The chip was formed by PMMA microspheres through a self-assembly method. As a good adsorbent of peptides, PMMA microspheres make peptides distributed uniformly on the chip, resulting in the elimination of the coffee ring effect and the homogeneous co-crystallization of peptides/matrix. The relative standard deviations (∼ 10 %) and coefficients of determination (∼ 0.99) are satisfactory when detecting 3 standard peptides, which show that the chip is applicable for rapid quantification of peptides. The limit of detection is improved by 1000-fold on this chip compared with conventional steel target because of the enrichment of the super-hydrophobicity. It can be concluded that the detection reproducibility and sensitivity are significantly improved by the super-hydrophobic structured-PMMA chip. The applicability of the chip for practical samples is demonstrated by quantifying bacitracin A in milk extract and bradykinin1-7 in serum with good linearities. The structured-PMMA chips exhibit competitive advantages for quantitative analysis of peptides, which have a great potential for proteomic research.
研究领域:
分析化学
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生物医学工程
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蛋白质组学
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DOI
高影响因子
TALANTA
·
2016
·
IF 4.035
Wenjie Gao,Henghui Li,Yanyan Liu,Yuhong Liu,Xiaojun Feng,Bi-Feng Liu,Xin Liu
Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) has become an important technology for glycan analysis due to its ease of operation, short analysis time and impurity tolerance. However, the low ionization efficiency of N-glycans led to the difficulty in analyzing glycans of low abundance in complex biological samples due to the lack of basic site for protonation. Therefore, highly sensitive method for the glycans analysis is in urgent demand. Here we report a new strategy to introduce a permanent charge at the reducing end of N-linked glycans by a one pot reaction , where glycosylamines that were obtained by rapid deglycosylation within 5 min were labeled with N-succinimidyloxycarbonylmethyl tris (2,4,6- trimethoxyphenyl) phosphonium bromide (TMPP-Ac-OSu). With TMPP-Ac labeling, more than 50 fold enhancement in the sensitivity of method was achieved for neutral glycans from ribonuclease B (RNase B) in comparison to their native counterparts. In combination with methylamidation of sialic acid residues, this novel developed strategy could also be used for sialylated glycans analysis from sialoglycoproteins and complex serum sample. As a result, more than 50 glycans were detected with only 25 nL human serum sample.
研究领域:
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
ANALYTICA CHIMICA ACTA
·
2017
·
IF 4.95
Chang Wang,Yike Wu,Liang Zhang,Bi-Feng Liu,Yawei Lin,Xin Liu
Quantitative analysis of glycans is an emerging field in glycomic research. Herein we present a rapid and effective dual-labeling strategy, in the combination of isotopic derivatization of N -glycosylamine-based glycans by d0/d5-benzoyl chloride and methylamidation of sialic acids , to relatively quantify both neutral and sialylated N -glycans simultaneously by MALDI-MS. The derivatization efficiencies were increased by microwave-accelerated deglycosylation which not only largely reduce the time of glycoprotein deglycosylation but also inhibit the hydrolysis of intermediate glycosylamines produced by PNGase F digestion. Three model glycoproteins, including RNase B, bovine fetuin and IgG from human serum, were applied to validate this technique. Results showed that the glycans from microgram level of glycoprotein can be successfully quantified with high reproducibility and the whole time of analytical procedure was shortened to 4 h. Furthermore, this proposed method was applied for the comparative analysis of N -glycans from serum of healthy donors and multiple myeloma patients. It was found that five N -glycans may be as the potential biomarkers for rapid detection of early multiple myeloma, indicating the feasibility of this strategy in monitoring subtle quantitative differences of serum glycomics.
研究领域:
生物化学
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医学诊断
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糖组学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
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2021
·
IF 3.205
Xiao-Pan Liu,Wen-Qian Sun,Tong-Xin Liu,Bing-Bing Liu,Chang-Po Chen
Due to the strong background interferences in the low-mass region and poor reproducibility of conventional organic matrices, it is of great importance to develop a novel matrix for matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) to qualitatively and quantitatively analyze small molecules. In this work, water-soluble fullerenol C 60 (OH) 24–26 was selected as a MALDI matrix for the analysis of low-molecular-weight compounds in consideration of optical absorption property, water solubility and stability. Compared with the traditional matrices, fullerenol demonstrated lower background interference and stronger peak intensity. In addition, the hydrophilic fullerenol could avoid the heterogeneous crystallization in sample preparation, increase the reproducibility and sensitivity of MALDI-MS, and ameliorate quantitative analysis of small molecules. With saccharin as model analyte, quantitative analysis was carried out using fullerenol as matrix. The results demonstrated satisfying reproducibility and good tolerance to salt. The limit-of-detection of the quantitative analysis was as low as 4 pmol, and the linear range is 1–100 μg mL −1 with R 2 greater than 0.99. The analytical results also showed excellent precision and accuracy, low matrix effect and good recovery rate. Fullerenol as a potential matrix was further validated in the quantification of saccharin sodium in different real food samples, such as nuts and drinks. This work not only confirms the potential of fullerenol for the quantitative analysis in food field, but also provides a new technique for rapid analysis of small molecules.
研究领域:
分析化学
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食品科学
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质谱技术
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2021
·
IF 6.057
Weiwei Tang,Andrew Gordon,Hui-ying Wang,Ping Li,Jun Chen,Bin Li
The development of in situ methods for the analysis and visualization of enzyme activity is of paramount importance in drug discovery, research, and development. In this work, the functionalized and array patterned indium tin oxide (ITO) glass slides were fabricated by non-covalent immobilization of amphipathic phospholipid-tagged peptides encompassing the thrombin cleavage site on steric acid-modified ITO slides. The fabricated peptide arrays provide 60 spots per slide, and are compatible with matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) measurement, free matrix peak interference, and tolerance to repeated aqueous washing. The peptide arrays were used for the investigation of thrombin activity and screening for its potential inhibitors. The thrombin activity and its Michaelis-Menten constant (K m ) for immobilized peptide substrate was determined using developed MALDI MS peptide array. To investigate the applicability and effectiveness of peptide arrays, the anti-thrombin activity of grape seed proanthocyanidins with different degrees of polymerization (DP) was monitored and visualized. MALDI MS imaging results showed that the fractions of proanthocyanidins with the mean DP of 4.61–6.82 had good thrombin inhibitory activity and their half-maximal inhibitory concentration (IC 50 ) were below 10 μg/mL. Therefore, the developed peptide array is a reliable platform for the discovery of natural thrombin inhibitors .
研究领域:
药物发现
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分析化学
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药学
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生物化学
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产品关联
DOI
PubMed
高影响因子
ANALYST
·
2021
·Vol. 146
·期 9
·2955-2964
·
IF 4.616
Jiewei Yang,Chenyu Wang,Li Luo,Zhi Li,Bin Xu,Lei Guo,Jianwei Xie
Herein, we report an improved MALDI-MS method for active ricin to contribute toward countermeasures against its real threat to the public. Compared with commonly used DNA or RNA substrates, the deoxynucleobase-hybrid oligonucleotide (RNA_dA, Rd) substrate containing functional GGA loop was revealed as a substrate with more potential and used for the first time in ricin measurement via MALDI-MS. The Rd sequence greatly prompted ricin to exhibit its catalytic activity as rRNA N-glycosylase in ex vitro condition, which was supported by molecular docking simulation and enzymatic parameters depicted in MALDI-MS. Furthermore, we discovered that a highly pure matrix was the most crucial parameter for enhancing the sensitivity, which addressed the major obstacle encountered in the oligo(deoxy)nucleotide measurement, i.e., the interfering alkali metal ion-adducted signals in MALDI-MS. After the optimization of pH and enzymatic reaction buffer composition in this ex vitro condition, this method can provide a wide linearity of up to three orders of magnitude, i.e., 1–5000 ng mL−1, and a high sensitivity of 1 ng mL−1 without any enrichment. Denatured and active ricin could be distinctly differentiated, and the application to practical samples from one international exercise and a soft drink proved the feasibility of this new method. We believe this MALDI-MS method can contribute to the first response to ricin occurrence events in public safety and security, as well as pave a new way for a deep understanding of ricin and other type II ribosome inactivating proteins involved toxicology.
研究领域:
分析化学
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毒理学
·
生物技术
·
公共卫生
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产品关联
DOI
PubMed
高影响因子
CHINESE JOURNAL OF ANALYTICAL CHEMISTRY
·
2017
·
IF 0.795
Xing-Li PEI,Yu-Yu HUANG,Can GONG,Xu XU
The ionic liquid butylamine 2,5-dihydroxybenzoate (DHB-BuN) was chosen as the matrix for improving reproducibility of oligosaccharides quantification, and imaging oligosaccharides in soybean and leaf by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS). In the experiment, ionic liquid matrix of methanol solution (20%, V/V ) was simply covered onto the sample slice, and MS imaging experiment was thus performed to analyze oligosaccharides and their distribution in soybean and soybean leaf in positive ion mode with laser power of 70%. As a result, the MALDI-MS analysis of three oligosaccharides including sucrose, raffinose and stachyose was performed using the ionic liquid matrix, and the reproducibility RSD was less than 3% for in-spot and less than 4% for spot-to-spot. The linear correlation coefficients ( R 2 ) of standard curves between MS peak intensity and oligosaccharides concentrations raging from 0.062 mg mL −1 to 1.00 mg mL −1 were all no less than 0.996, which showed a great potential in quantitative analysis of oligosaccharides by MALDI-MS. The matrix was then used for the imaging analysis of disaccharide, trisaccharide and tetrasaccharide on the surfaces of soybean chip and soybean leaf by MALDI-MS, and the mass spectrometric imaging of the three kinds of oligosaccharides was obtained with a spatial resolution of 150 μm. The result indicated that the distribution of three kinds of oligosaccharides in the soybean was roughly homogeneous, and mostly at its tip part in the leaf. The contents of the three oligosaccharides could be estimated by their MS peak intensity and above standard curve. This study suggested that the ionic liquid DHB-BuN matrix could be well applied in MALDI-MS imaging analysis of oligosaccharides in different samples.
研究领域:
分析化学
·
植物学
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产品关联
DOI
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2024
·
IF 3.8
Sun Zhi,Wang Fangfang,Li Wenxuan,Ren Ruobing,Zhou Peipei,Jia Qingquan,Zhao Lingguo,Chen Di,Zuo Lihua
Therapeutic drug monitoring is essential for ensuring the efficacy and safety of medications. This study introduces a streamlined approach that combines pipette-tip solid-phase extraction (PT-SPE) with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), facilitating rapid and high-throughput monitoring of drug concentrations. As a demonstration, this method was applied to the extraction and quantification of antidepressants in serum. Utilizing Zip-Tip C18, the method enabled the extraction of antidepressants from complex biological matrices in less than 2 min, with the subsequent MALDI-MS analysis yielding results in just 1 min. Optimal extraction recoveries were achieved using a sampling solution at pH 9.0 and a 10 μL ethanol desorption solution containing 0.1% phosphoric acid. For MALDI analysis, 2,5-dihydroxybenzoic acid was identified as the most effective matrix for producing the highest signal intensity. The quantification strategy exhibited robust linearities ( R 2 ≥ 0.997) and satisfactory limits of quantification, ranging from 0.05 to 0.5 μg/mL for a suite of antidepressants. The application for monitoring dynamic concentration changes of antidepressants in rat serum emphasized the method’s efficacy. This strategy offers the advantages of high throughput, minimal sample usage, environmental sustainability, and simplicity, providing ideas and a reference basis for the subsequent development of methods for therapeutic drug monitoring. Graphical Abstract
研究领域:
药物分析
·
临床医学
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产品关联
DOI
PubMed
高影响因子
RSC Advances
·
2017
·Vol. 7
·期 57
·35687-35693
·
IF 3.108
Wenjie Gao,Yanhua Jiang,Zhihui Zhang,Yifang Zhang,Yanyan Liu,Yanhong Zhou,Xin Liu
Conventional protocols for cellular N-glycan profiling often need several time-consuming and labor-intensive steps, and a large amount of material (5–20 × 106 cells) is required. In this study, an optimized co-derivatization method was applied for convenient, rapid and highly-sensitive analysis of cellular N-glycan. Taking human cervical carcinoma cells (HeLa) as a model, the required amount for comprehensive cellular N-glycans analysis was reduced down to an original cell number of 105 and the total analysis time was decreased from several days to several hours. Good reproducibility of the method was also obtained with the CV averaging to less than 13.1%. In addition, compared to permethylation derivatization, more than 5-fold sensitivity improvement was achieved and more concise and clear mass spectra were obtained with the new developed method. As a preliminary study, this method was also successfully applied to reveal the difference in cellular N-glycan profiling of two heterogeneous live cell lines, human normal liver cells (L-02) and human hepatocyte carcinoma cells (HepG2), showing great potential for high-throughput analysis of N-glycans from limited cell samples, such as primary cell lines, cells obtained from cell sorting and small size cancer specimens.
研究领域:
生物化学
·
分析化学
·
医学研究
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产品关联
DOI
高影响因子
JOURNAL OF MASS SPECTROMETRY
·
2024
·Vol. 59
·期 7
·e5058
·
IF 1.9
Xintong Zhao,Juan Ren,Ze Wang,Xiangfeng Chen
Analysis of noncovalent interactions between natural products and proteins is important for rapid screening of active ingredients and understanding their pharmacological activities. In this work, the intensity fading MALDI-TOF mass spectrometry (IF-MALDI-MS) method with improved reproducibility was implemented to investigate the binding interactions between saponins from Panax notoginseng and lysozyme. The benchmark IF-MALDI-MS experiment was established using N , N ′, N ″-triacetylchitotriose-lysozyme as a model system. The reproducibility of ion intensities in IF-MALDI-MS was improved by scanning the whole sample deposition with a focused laser beam. The relative standard deviation (RSD) of deposition scanning IF-MALDI-MS is 5.7%. Similar decay trends of the relative intensities of notoginseng saponins against increasing amounts of lysozyme were observed for all six notoginseng saponins. The half-maximal fading concentration (FC 50 ) was calculated to quantitatively characterize the binding affinity of each ligand based on the decay curve. According to the FC 50 values obtained, the binding affinities of the six notoginseng saponins were evaluated in the following order: notoginsenoside S > notoginsenoside Fc > ginsenoside Rb1 > ginsenoside Rd > notoginsenoside Ft1 > ginsenoside Rg1. The binding order was in accordance with molecular docking studies, which showed hydrogen bonding might play a key role in stabilizing the binding interaction. Our results demonstrated that deposition scanning IF-MALDI-MS can provide valuable information on the noncovalent interactions between ligands and proteins.
研究领域:
分析化学
·
药理学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）