JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2025
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IF 6.2
Xin-Ze Wu,Kai-Wen Tan,Xu Zhang,Jun-Jie Tan,Na An,Fang Cong,Wenbin Wu,Azamat Temerdashev,Quan-Fei Zhu,Hong Wang,Yu-Qi Feng
Fatty acid esters of hydroxy fatty acids (FAHFAs) are emerging bioactive lipids in edible oils, but their trace-level presence and matrix complexity hinder accurate analysis. Here, we designed a novel chemoselective probe piperazine-modified silica (SiO2-PP) that specifically captures FAHFAs while removing almost totally triglycerides in a one-step labeling reaction. The probe enabled single-batch sample pretreatment within 30 min, integrating capture, purification, and signal amplification. Coupled with liquid chromatography–mass spectrometry, the method achieved high sensitivity (LOD: 0.0039–0.033 ng/mL) and enabled quantification of 47 FAHFA isomers in 47 vegetable oil samples, with concentrations ranging from 0.56 to 1.76 × 104 ng/g. Five dominant isomers (e.g., 10-OAHSA) were identified, with olive and peanut oils exhibiting the highest FAHFA concentrations. This approach provides a promising platform for high-throughput functional lipid profiling, effectively overcoming the inefficiencies of traditional methods while significantly enhancing the assessment of vegetable oil quality.
研究领域:
分析化学
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脂质组学
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食品科学
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生物活性脂质
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质谱分析
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高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
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2021
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IF 3.978
Mianxing Luo,Meng Zhang,Guo Chen,Jun Zhao,Hongwei Guo
Functional carrier capable of covalently binding proteins from crude lysate under mild condition has become increasingly important in bioprocess. In this paper, a magnetic microbead that can covalently immobilize specific proteins directly from crude protein solution was developed, and its application conditions and stability were evaluated. The SpyCatcher was firstly modified with N-terminal cysteine by genetic methods and then the cys-SpyCatcher was covalently immobilized on epoxy-modified magnetic beads by epoxy-thiol reaction. The reaction between the epoxy and the only terminal thiol group in cys-SpyCatcher ensured the same orientation of the cys-SpyCatcher on the particle surface, while the maximum binding capacity of cys-SpyCatcher reached 23.6 μg/mg under the condition of 250 mM Na 2 SO 4 , pH 8.5 at initial cys-SpyCatcher concentration of 3 mg/mL. The functional magnetic microsphere ( [email protected] ) showed covalent site-specific capture property for enhanced green fluorescent protein fused with SpyTag (SpyTag-EGFP). A maximum binding capacity of (25 μg SpyTag-EGFP)/(mg beads) was far more than that of the previous carrier with SpyCatcher. In addition, the [email protected] could specifically capture the SpyTagged protein directly from the cell lysate, and this carrier showed good stability at pH 4–9 and temperature of 30–70ºC. Therefore, as a universal covalent capture carrier, it is believed that the [email protected] is useful in one-step purification and site-specific immobilization of the SpyTagged protein in bioprocess or biomedical application.
研究领域:
生物技术
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生物化学
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蛋白质工程
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DOI
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
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2025
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IF 11.3
Ruidong Zhou,Xinyuan Zhang,Yupeng Wen,Wendi Huo,Yingchun Mu,Huan Liu,Chaoying Zhang,Jincheng Li
Multi-walled carbon nanotubes (MWCNTs) are known for their excellent purification properties in QuEChERS-based pesticide analysis, but their potential release and non-recyclability pose environmental and economic challenges. In this study, a novel strategy is proposed to immobilize MWCNTs with chitosan, forming the MWCNTs@chitosan composite, which enables the recyclability of MWCNTs. This composite was employed in a one-step pressure filtration purification method to simultaneously purify extracts of 49 pesticides from aquatic products. Coupled with HPLC-MS/MS, an optimized QuEChERS method was successfully validated for the determination of 49 pesticides. The MWCNTs@chitosan composite demonstrated excellent clean-up capability, effectively removing interfering substances from crude extracts while maintaining recoveries of 71.2–119.4 % and relative standard deviations (RSDs) below 20.0 %. These results demonstrate the composite’s suitability as a QuEChERS cleanup material, with potential applications in other complex matrices.
研究领域:
环境科学
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分析化学
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食品安全
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纳米技术
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材料科学
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JOURNAL OF CHROMATOGRAPHY A
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2022
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IF 4.601
Jun Ren,Hao Xiong,Chundong Huang,Fangling Ji,Lingyun Jia
Camelid-derived nanobody is emerging as a resourceful platform for developing immunoaffinity ligands for chromatography applications. Featured by high affinity and selectivity, BC2 nanobody (BC2-Nb), which can recognize a specific epitope tag (PDRKAAVSHWQQ, termed BC2T), is potential to be developed as a general tool for recombinant protein purification. However, excessively high affinity between binding partners makes the desorption of products less efficient and limits its application. Aiming to improve elution efficiency, structure-guided mutations of BC2-Nb were conducted to adjust the structural flexibility of its antigen-binding site. Six ligand variants were obtained with their binding affinity decreasing by about 100-fold. Among them, one mutated BC2-Nb named 44D was chosen to prepare immunoaffinity resin, and its adsorption and elution performance were well characterized. The site-directed mutation led to the equilibrium dissociation constant (K D ) of BC2-Nb changing from 1.4 × 10 −9 M to 1.4 × 10 −7 M (44D). The resin using 44D as ligand retained a static binding capacity of 19.14 mg/mL toward BC2T-fused enhanced green fluorescent protein (eGFP-BC2T). Significantly improved elution efficiency was obtained with the mutated ligand. Protein recovery reached 94% at pH 3.5 for 44D-based resin, while the resin based on original BC2-Nb could only achieve its highest recovery of 84% at pH 2. In addition, a neutral elution condition (1 M arginine containing 50% propylene glycol, pH 7.4) was also found effective, which allowed a product recovery of 95%. The resin enabled direct capturing of eGFP-BC2T from bacterial lysates, and the one-step purification with the both elution conditions could achieve a product purity of more than 90%. This study provided a promising affinity ligand, and also proved the feasibility of controlling the elution process of nanobody-based affinity resin through the strategy of binding sites modification.
研究领域:
生物技术
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蛋白质工程
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色谱
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生物化学
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PubMed
高影响因子
危险品化学品经营许可证（带存储）