JOURNAL OF CHROMATOGRAPHY A
·
2021
·
IF 4.759
Canhong Zhu,Jiani Wu,Xueting Jin,Yinghua Yan,Chuan-Fan Ding,Keqi Tang,Di Zhang
Chemical modification of biological materials is indispensable for enrichment of phosphorylated peptides. In this work, we synthesized a biomimetic honeycombed affinity chromatography (IMAC) adsorbent by preparing Crosslinked Chitosan, chelating aminomethyl phosphate decorated with Ti (IV) cation. The as-prepared CTSM@AMPA-Ti 4+ composites with stable structure, low steric hindrance, and high Ti 4+ loading amount were used as a promising adsorbent for enrichment of phosphopeptides. CTSM@AMPA-Ti 4+ showed extremely high sensitivity (0.4 fmol) and selectivity at a low composition molar ratio of β-casein/BSA (1:1000). What's more, it can keep its performance in the case that used to capture phosphorylated peptides from standard protein ten times or soaking in the acid/base solution for a long time. In addition, CTSM@AMPA-Ti 4+ successfully captured 35 phosphorylated peptides from human saliva. This study offers a way about diversiform functionalization of CTSM in phosphoproteome analysis and disease research.
研究领域:
生物化学
·
分析化学
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DOI
高影响因子
BIOCHEMICAL PHARMACOLOGY
·
2025
·
IF 5.6
Qianqian Wang,Xin Yuan,Cuijuan Liu,Ying Huang,Lin Li,Yimin Zhu
CAR-T cell therapy has been proven to be effective on hematological tumors, although graft-versus-host disease and cytokine release syndrome(CRS) limit its application to a certain extent. However, CAR-T therapy for solid tumors met challenges, among which the lack of tumor-specific antigens (TSA) and immunosuppressive tumor microenvironment (TME) are the most important factors. CAR-NK could be a good alternative to CAR-T in some ways since they can induce mild CRS and are independent of HLA-matching, but the efficacy of CAR-NKs remains limited in solid tumors. CAR cells armed with multiple tumor targeting molecules may obtain higher therapeutic efficacy against solid tumors. Due to large molecular weight, multivalent scFvs cannot be displayed efficiently on T cells and the high affinity of scFv to the target makes it easy to cause on-target, off-tumor(OTOT) toxicity. Peptides with low molecular weight and slightly lower affinity than scFvs allow immune cells to display multiple peptides to increase killing ability and reduce OTOT toxicity. In our study, peptide-based CAR-NK cells were designed to solve the dilemma of CAR-T in solid tumors. Firstly, the peptide-based CAR-NK92MI cells with A1 peptide were constructed and their inhibitory effects on the growth of A549 tumor cells were identified. Secondly, the tri-specific CAR-NK92MI cells with peptides that simultaneously targeted PD-L1, EGFR and VEGFR2 were developed for the combinatory therapy. Tri-specific CAR-NK92MI exhibited comparable killing activities to scFv-based CAR-NK92MI. Moreover, peptide-based CAR NK92MI mitigated OTOT toxicity. Our study implied that peptide-based CAR-NKs could behave as promising tools in solid tumor
研究领域:
免疫学
·
肿瘤学
·
细胞治疗
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 8
·1-8
·
IF 5.479
Jiang Dandan,Li Zheng,Jia Qiong
The design and preparation of magnetic cucurbit[6]uril hypercrosslinked
with polymers are described. The materials have a large specific surface, abundant
mesopores and cavities, and display superparamagnetism. They were applied to the
enrichment of ubiquitinated peptides from standard protein digests. Following
desorption with 0.15% TFA, the peptides were quantified by MALDI-TOF MS. The method
has a detection limit of 2 fmol·μL −1 and a mass
ratio selectivity of 1:5000 as shown for ubiquitin and bovine serum albumin. The
materials enable selective capture of ubiquitinated peptides from genuine samples
comprising of oyster mushroom and human serum. This demonstrates their potential for
the analysis of low-level ubiquitin in complex samples.
Schematic presentation for the synthesis of magnetic
cucurbit[6]urils hypercrosslinked polymers
(MagCB[6]-HCPs).
研究领域:
生物化学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF FLUORESCENCE
·
2013
·Vol. 24
·期 2
·533-539
·
IF 1.789
Liu Chengxuan,Liang Gaiting,Liu Zhen,Zu Lily
Time-resolved fluorescence anisotropy spectroscopy was applied to study the interaction between a peptide truncated from the binding site of tumor suppressor p53 protein and the DNAs covalently labeled with 6-carboxyfluorescein (FAM) dye. Fluorescence intensity quenching and changes of anisotropy decay lifetime were monitored when FAM labeled DNA formed complex with the peptide. The results demonstrated that the sequence of DNA could not define the binding specificity between the peptide and DNA. But the anisotropy decay of FAM can be used to examine the binding affinity of the peptide to DNA. The fluorescent dynamics of FAM can also be used to represent the rigidity of the complex formed between the peptide and DNA.
研究领域:
生物化学
·
分子生物学
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DOI
PubMed
高影响因子
Materials Science & Engineering C-Materials for Biological Applications
·
2020
·
IF 5.88
Peichuang Li,Xiaojing Li,Wanhao Cai,Huiqing Chen,Hang Chen,Rui Wang,Yuancong Zhao,Jin Wang,Nan Huang
As an important class of biomaterials，bionics inspired materials has been widely used in creating extracorporeal and implantable medical devices. However, specific service environment is often faced with multiple requirements rather than single function. Herein, we designed a phospholipid-based multifunctional coating with phospholipids-based polymers, type I collagen (Col-I) and Arg-Glu-Asp-Val (REDV) peptide, via layer-by-layer assembly. The successful synthesis of the polymers and the coating is proved by a series of characterization methods including Fourier transforming infrared spectra (FTIR), proton nuclear magnetic resonance ( 1 H NMR), ultraviolet-visible spectra (UV) and X-ray photoelectron spectroscopy (XPS), while the assembly process and quality change of the coating were monitored via quartz crystal microbalance (QCM). Besides, hydrophilicity and roughness of this coating was analyzed via water contact angle (WCA) and atomic force microscope (AFM), respectively. Finally, results from platelet adhesion, activation assay, smooth muscle cells (SMCs) and endothelial cells (ECs) cultures indicated that the multifunctional coating could strongly inhibit platelet adhesion and SMCs proliferation, hence provide practical application of the coating with good biocompatibility, especially the anticoagulant property and cell compatibility. It is expected that this coating may be used in blood-contacting fields such as cardiovascular stent or other devices in the future.
研究领域:
生物材料学
·
生物医药工程
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DOI
PubMed
高影响因子
FOOD RESEARCH INTERNATIONAL
·
2024
·
IF 7
Xialin Chen,Dandan Li,Rongrong He,Jiajia Wang,Ru Yang,Mengyu Qian,Liang Cao,Yongwen Zhang,Zhenzhong Wang,Wei Xiao
Polyrhachis dives ( P. dives ) is a traditional edible insect with high nutritional value and is the only ant species that is administered as medicine and food. Therefore, it is of great significance to identify the authenticity of the species. In the current study, peptidomics analysis combining untargeted MS with targeted MS was developed and applied for the identification of specific peptides in P. dives and the establishment of quality control. A total of 16 specific peptides were screened, and 7 specific peptides with satisfactory signal response were obtained by targeted mass spectrometry, which could be used to distinguish P. dives from other ant species and ensure the quality of edible ants. Subsequently, the 7 specific peptides were quantitatively analyzed, followed by clustering analysis and biological activity experiments to categorize them into distinct qualities, providing valuable references for further development of quality control methods for P. dives .
研究领域:
昆虫学
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食品科学
·
质谱分析
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DOI
PubMed
高影响因子
FOOD HYDROCOLLOIDS
·
2025
·
IF 12.4
Le-Ying Yang,Zong-Cai Tu,Hui Wang,Xue-Ke Sun,Yu-Ting Hu,Yu Chen,Zi-Zi Hu,Xiao-Mei Sha
The physical properties and gelatin identification of gummy candies could be affected in the storage process by acid hydrolysis and glycation reaction. In this work, the physical properties including texture, color, appearance and molecular weight distribution were evaluated in the porcine gelatin-containing gummy candies that were stored under the accelerated model at different storage stages. In addition, the characteristic peptides of porcine gelatin were identified by high performance liquid chromatography and linear ion trap/Orbitrap MS/MS (HPLC-LTQ/Orbitrap MS/MS) to investigate their changes. The results showed that hydrolysis was the main reaction in the early stage of storage. The hardness, gumminess, chewiness and high molecular weight peptides content of gummy candies decreased greatly with the extension of storage time. Free amino groups were produced by gelatin hydrolysis to promote glycation and glycosidic bond. In the later stage of storage, glycation was dominant, which led to the decrease of low molecular weight peptides. Moreover, HPLC-MS/MS identified 275, 260, 243, 248 and 257 characteristic peptides of porcine gelatin from gummy candies treated by the accelerated model in 0, 14, 28, 42 and 56 days, respectively. Among them, 130 characteristic peptides were found to remain unchanged during the shelf-life of gummy candies. By further comparison with the identification under various food processing conditions, 6 stable characteristic peptides were reserved until the end. They could provide the basis for the subsequent traceability of porcine gelatin.
研究领域:
食品科学
·
生物化学
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产品关联
DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2024
·
IF 5.7
Yuyan Wei,Ling Xie,Bertrand Muhoza,Qian Liu,Shiqing Song
The present study investigated the impact of four chicken liver protein hydrolysate-based cat food attractants on palatability. Aroma compounds were analyzed in these attractants, which were subsequently sprayed onto four different types of cat foods. Results revealed that CF4 exhibited the highest intake ratio and the first choice ratio, followed by CF2 sample. Orthogonal partial least-squares discriminant analysis (OPLS-DA) demonstrated significant differences among 50 volatile compounds identified from the four cat foods. Using variable importance in projection (VIP) values, we selected 17 key flavor compounds responsible for distinguishing between the four cat foods. Peptides with a molecular mass <180 Da showed correlation with nonanoic acid and cedrol, while those >3000 Da correlated with hexanoic acid ethyl ester. Regression coefficients (RCs) calculated from partial least-squares regression (PLSR) results showed positive correlations between compound content and palatability for six compounds, whereas negative correlations were observed for ten compounds. Validation experiments confirmed that nonanal, 2-propylpyridine, and 3-octen-2-one enhanced palatability and correlated with peptides ranging from 180 to 500 Da; conversely, nonanoic acid ethyl ester and 3-methyl-pentanoic acid reduced palatability and correlated with peptides ranging from 1000 to 3000 Da.
研究领域:
食品科学
·
动物营养学
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产品关联
DOI
PubMed
高影响因子
Nanoscale
·
2013
·Vol. 5
·期 22
·10936-10944
·
IF 6.233
Hao Wan,Hongqiang Qin,Zhichao Xiong,Weibing Zhang,Hanfa Zou
Magnetic mesoporous carbon microspheres with a yolk–shell structure (YSMMCS) have been prepared via a new in situ carbon source strategy. The material was fabricated by two shells coated onto the Fe3O4 particles; the inner dense and thick silica shell could protect the magnetic core from harsh acidic solvents as well as induce the void between the core and the outer shell for the yolk–shell structure, while the outer organosilica shell was used as the template and carbon source for in situ preparation of a carbon shell with mesoporous structure. A C18-alkyl chain was incorporated in situ as the carbon precursor efficiently, avoiding the conventional infiltration step, which was very difficult to manipulate and time-consuming with the possibility of losing the carbon precursor. The resulting yolk–shell magnetic mesoporous carbon microspheres exhibited a high surface area (273.15 m2 g−1), a large pore volume (0.31 cm3 g−1), and a strong magnetic response (a saturation magnetization value of 34.57 emu g−1). As a result of the void between the core and the outer shell and the π–π stacking effect, adsorption capacity reached 191.64 mg g−1 by using Rhodamine B as a standard analyte, indicating the great potential application of the material as drug carriers. Owing to the inherent hydrophobicity and high surface area, the composite material showed better performance in the enrichment of peptides than a magnetic mesoporous silica material (Fe2O3@nSiO2@mSiO2). According to the LC-MS/MS results, about 51 and 29 nonredundant peptides were identified from tryptic digests of 5 nM BSA. Additionally, taking advantage of the mesoporous structure and strong magnetic response, the material was utilized to selectively extract low abundance endogenous peptides from human serum in the presence of high abundance proteins. Based on the LC-MS/MS results, 962 endogenous peptides were obtained by 2.5 mg YSMMCS relative to 539 endogenous peptides by 5 mg Fe2O3@nSiO2@mSiO2, confirming the outstanding performance of YSMMCS in peptidome analysis.
研究领域:
材料科学
·
生物医学
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DOI
PubMed
高影响因子
MARINE BIOTECHNOLOGY
·
2025
·Vol. 27
·期 2
·1-18
·
IF 2.8
Li Xue,Meng Fanying,Sun Tong,Hao Zezhuang,Wang Yihua,Jiang Yunfei,Wang Ye,Li Yong,Ding Yuling
This article aims to reveal the optimal peptide segment with antioxidant activity from Dalian Stichopus japonicus , investigate its anti-melanogenesis effect, and elucidate its mechanisms of action both in vitro and in vivo. The best antioxidant alcalase hydrolysates, identified by the previous screening of proteases, was isolated by ultrafiltration; it was found that the components with a molecular weight of ≤ 3 kDa exhibited the best activity. The chemical components were characterized using LC–MS/MS. Through molecular docking, GPIGF was identified as the peptide segment with the best antioxidant and melanogenesis-inhibitory activity. A search in the NCBI database revealed that GPIGF is a newly obtained natural oligopeptide. Further experiments with synthesized GPIGF in vitro showed that it effectively reduces cell apoptosis and damage, and inhibits the expression of melanin-related genes, including tyrosinase (TYR), and associated proteins TRP-1, TRP-2, and MITF. In vivo experiments with the zebrafish model demonstrated that GPIGF significantly inhibits AAPH-induced apoptosis in zebrafish larvae, reduces the production of ROS, and suppresses melanin generation on the skin surface without exhibiting embryotoxicity. This study provides a research foundation for the development of antioxidants from Dalian Stichopus japonicus , which could serve as natural whitening and anti-aging agents, supporting its integrated utilization and development.
研究领域:
生物化学
·
药理学
·
海洋生物学
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产品关联
DOI
PubMed
高影响因子
Nature Protocols
·
2013
·Vol. 8
·期 3
·461-480
·
IF 7.96
Zhou Houjiang,Ye Mingliang,Dong Jing,Corradini Eleonora,Cristobal Alba,Heck Albert J R,Zou Hanfa,Mohammed Shabaz
Mass spectrometry (MS)-based proteomics has become the preferred tool for the analysis of protein phosphorylation. To be successful at such an endeavor, there is a requirement for an efficient enrichment of phosphopeptides. This is necessary because of the substoichiometric nature of phosphorylation at a given site and the complexity of the cell. Recently, new alternative materials have emerged that allow excellent and robust enrichment of phosphopeptides. These monodisperse microsphere–based immobilized metal ion affinity chromatography (IMAC) resins incorporate a flexible linker terminated with phosphonate groups that chelate either zirconium or titanium ions. The chelated zirconium or titanium ions bind specifically to phosphopeptides, with an affinity that is similar to that of other widely used metal oxide affinity chromatography materials (typically TiO 2 ). Here we present a detailed protocol for the preparation of monodisperse microsphere–based Ti 4+ -IMAC adsorbents and the subsequent enrichment process. Furthermore, we discuss general pitfalls and crucial steps in the preparation of phosphoproteomics samples before enrichment and, just as importantly, in the subsequent mass spectrometric analysis. Key points such as lysis, preparation of the chromatographic system for analysis and the most appropriate methods for sequencing phosphopeptides are discussed. Bioinformatics analysis specifically relating to site localization is also addressed. Finally, we demonstrate how the protocols provided are appropriate for both single-protein analysis and the screening of entire phosphoproteomes. It takes ∼ 2 weeks to complete the protocol: 1 week to prepare the Ti 4+ -IMAC material, 2 d for sample preparation, 3 d for MS analysis of the enriched sample and 2 d for data analysis.
研究领域:
蛋白质组学
·
生物化学
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2017
·
IF 4.162
Linge Nian,Yue Hu,Caihong Fu,Chen Song,Jie Wang,Jianxi Xiao
The development of novel assays to detect collagen fragments is of utmost importance for diagnostic, prognostic and therapeutic decisions in various collagen-related diseases, and one essential question is to discover probe peptides that can specifically recognize target collagen sequences. Herein we have developed the fluorescence self-quenching assay as a convenient tool to screen the capability of a series of fluorescent probe peptides of variable lengths to bind with target collagen peptides. We have revealed that the targeting ability of probe peptides is length-dependent, and have discovered a relatively short probe peptide FAM-G(POG) 8 capable to identify the target peptide. We have further demonstrated that fluorescence self-quenching assay together with this short probe peptide can be applied to specifically detect the desired collagen fragment in complex biological media. Fluorescence self-quenching assay provides a powerful new tool to discover effective peptides for the recognition of collagen biomarkers, and it may have great potential to identify probe peptides for various protein biomarkers involved in pathological conditions.
研究领域:
生物化学
·
分子生物学
·
诊断学
✓ 已验证
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）