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SENSORS AND ACTUATORS B-CHEMICAL · 2020 · IF 7.1

A sensitive lateral flow immunochromatographic strip with prussian blue nanoparticles mediated signal generation and cascade amplification

Meiling Tian,Wenyue Xie,Ting Zhang,Yang Liu,Zhisong Lu,Chang Ming Li,Yingshuai Liu

This work reports a low-cost, fast and highly sensitive lateral flow immunoassay (LFIA) platform with Prussian blue nanoparticle (PBNP) as a new reporter, which efficiently mediates an on-demand cascade signal amplification by catalyzing oxidation of chromogenic reagent 3,3′,5,5′-tetramethylbenzidine (TMB). In this case, PBNPs are accumulated on the test line through the specific immune interaction, resulting in a blue-colored line. The colorimetric signal can be further amplified via the peroxidase mimic PBNP-catalyzed TMB oxidation to produce an insoluble blue-violet precipitate. Under an optimal condition, a limit of detection (LOD) at 10 pg mL −1 is achieved for rabbit IgG (RIgG) detection, which is 2–3 orders of magnitude better than the conventional LFIA strip using colloidal gold nanoparticle as a label. A macromolecular RIgG and a small molecular mycotoxin ochratoxin A (OTA) are also successfully analyzed in mimicking samples, demonstrating its good practicability of the proposed platform in a variety of fields.

研究领域: 纳米技术 · 分析化学 · 生物传感器 · 免疫学
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ANALYST · 2013 ·Vol. 138 ·期 9 ·2624-2631 · IF 3.969

A paper-based surface-enhanced resonance Raman spectroscopic (SERRS) immunoassay using magnetic separation and enzyme-catalyzed reaction

Yuanyuan Chen,Hanwen Cheng,Kha Tram,Shengfeng Zhang,Yanhua Zhao,Liyang Han,Zengping Chen,Shuangyan Huan

In this study, a novel paper-based SERRS immunoassay based on magnetic separation and alkaline phosphatase (ALP) enzyme catalyzed hydrolysis reaction was developed. By using modified antibodies conjugated to magnetic beads, capture of mouse IgG followed by addition of ALP-labeled antibodies would form a sandwich-like immunoconjugate. After magnetic separation, 5-bromo-4-chloro-3-indolyl phosphate (BCIP), a low SERRS active compound, was added as the substrate for ALP to generate a high SERRS response. Detection was conducted on a silver colloid/PVP/filter paper SERS substrate by spotting a pre-aggregated silver colloid sol onto polyvinyl pyrrolidone (PVP) modified filter paper using a semi-automatic TLC sample applicator. The optimization of the highly SERS active paper-based substrate, dynamic hydrolysis process of BCIP, quantitative detection of IgG, and selectivity of the assay was studied in detail. By taking advantage of magnetic separation in order to decrease the background interference, the selective enzyme reaction involved in producing a highly SERRS active product could detect mouse IgG from 1 to 500 ng mL−1 with a LOD of 0.33 ng mL−1.

CAS / MDL: 6066-82-6
研究领域: 分析化学 · 生物传感器 · 免疫分析 · 光谱学
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ANALYTICA CHIMICA ACTA · 2021 · IF 6.558

Fluorescence detection of Staphylococcus aureus using vancomycin functionalized magnetic beads combined with rolling circle amplification in fruit juice

Yutong Wang,Zhengzheng Wang,Zhongxu Zhan,Ju Liu,Tingting Deng,Hengyi Xu

Staphylococcus aureus is a common foodborne pathogen that can cause a suppurative infection after eating contaminated food. Detection of S. aureus plays an important role in the food industry. In this study, a strategy for the detection of S. aureus using magnetic separation (MS) technology combined with rolling circle amplification (MS-RCA) was proposed. The strategy used antibiotics to capture bacteria and employed RCA products as signal output probes. Vancomycin (Van), as a commonly used antibiotic, can recognize peptidoglycan on the cell wall of Gram-positive bacteria and can effectively identify target bacteria. Therefore, we prepared BSAylated-Van functionalized magnetic beads (Van-MBs) for the pre-enrichment of S. aureus . To ensure the selectivity of this method, we used biotin-pig IgG to bind S. aureus . In addition, to amplify the output signal of the MS-RCA strategy, we introduced streptavidin (SA) and successfully obtained the [email protected] S. aureus @biotin-pig [email protected] @biotin-RCA probe complex and used the biotin-avidin-system (BAS) by combining magnetic separation technology and RCA technology to realize the enrichment and specific detection of S. aureus . Furthermore, by optimizing the experimental conditions such as the magnetic separation time and the amount of Van-MBs, the detection performance of this method was improved. Under the optimal conditions, the detection limit of this method for S. aureus was 3.3 × 10 2  CFU/mL in fruit juice, and it was less affected by other bacteria.

研究领域: 食品科学 · 微生物学 · 生物技术
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TOXICOLOGY · 2021 · IF 4.221

Exposure to a combination of MWCNTs and DBP causes splenic toxicity in mice

Tingting Zhou,Yueyan He,Yujie Qin,Bing Wang,Hongmao Zhang,Shumao Ding

The large conjugated π bond in the molecular structure of carbon nanotubes (CNTs) interacts with the benzene ring structure in di (n-butyl) phthalates (DBP) through a π - π bond . Compounds of CNTs and DBP form easily, becoming another environmental pollutant of concern. We explore whether CNTs entering animals slow down the degradation of the DBP adsorbed in the CNT cavity, thereby prolonging the "hormonal activity" of DBP. In our study, male BALb/c mice were used as experimental subjects divided into four groups: the control group; the multi-walled carbon nanotubes (MWCNTs) exposure group (10mg/kg/d); the DBP exposure group (2.15 mg/kg/d); and the compound exposure group (MWCNTs + DBP). After 30 days of exposure, the mice were sacrificed and their spleens used for immunotoxicology study. The results showed that the exposure groups exhibited splenomegaly and suffered severe oxidative damage to the spleen. In the compound exposure group: levels of IgA and IgG in the serum of the mice changed, and were significantly different from levels in both the MWCNTs and DBP exposure groups (p <0.05); the pathological sections of the spleen showed that the boundary between the white pulp area (WP) and the red pulp area (RP) was blurred, that the cell arrangement was loose, and that more red blood cells were retained in the spleen. Proteomics mass spectrometry analysis showed that compared with the control group, 70 proteins were up-regulated and 27 proteins were down-regulated in the MWCNTs group, 36 proteins were up-regulated and 23 proteins were down-regulated in the DBP group, 87 proteins were up-regulated and 21 proteins were down-regulated in the compound exposure group. The results of GO enrichment analysis and KEGG enrichment analysis of the differentially expressed proteins showed that the compound exposure harmed the spleen antigen recognition, processing, and presentation, inhibited the activation and proliferation of B cells and T cells, and hindered the adaptive immune responses. Our results showed that MWCNTs and DBP compounds can damage the spleen, and impair the innate and adaptive immune functions of the body.

研究领域: 环境毒理学 · 免疫学 · 蛋白质组学 · 分子生物学
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Surfaces and Interfaces · 2025 · IF 6.3

High-affinity capture of recombinant streptococcal protein G and Its application in serum proteomics with immunoglobulin G-coated magnetic nanoparticles

Dajun Zhang,Yufei Ma,Ying Wang,Xiangyu Liu,Mengfan Zhao,Yuanke Wang,Song Gao,Fu Ren,Qing Chen

A new solid-phase extraction agent, IgG@ Fe 3 O 4 -PEI, was successfully developed by conjugating immunoglobulin G (IgG) with amino-functionalized magnetic nanoparticles (Fe 3 O 4 -PEI). The IgG@Fe 3 O 4 -PEI composite was characterized using various techniques including infrared spectroscopy, Raman spectroscopy, magnetic property testing, Zeta potential measurements, energy spectrum analysis, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Because of the presence of a specific binding domain in the Fc fragment of IgG, the IgG@Fe 3 O 4 -PEI composite demonstrated highly selective adsorption of recombinant streptococcal G protein (rSPG) under optimal experimental conditions. At pH 6.0, the IgG@Fe 3 O 4 -PEI composite (0.5 mg) achieved a maximum adsorption efficiency of 93.96 % for rSPG at a concentration of 100 μg mL -1 . Adsorption model simulations indicated that the adsorption behavior followed the Langmuir adsorption model, with a theoretical maximum adsorption capacity of 125.0 mg g -1 . The enriched rSPG was eluted and recovered using Britton-Robinson (BR) buffer at pH 10.0, achieving a recovery efficiency of 95.8 %. In practical applications, the IgG@Fe 3 O 4 -PEI magnetic nanoparticles effectively enriched rSPG from an Escherichia coli expression system, and SDS-PAGE analysis confirmed the successful isolation of highly pure rSPG. Furthermore, the IgG@Fe 3 O 4 -PEI magnetic nanoparticles were used to capture proteins from human serum samples. After enrichment and processing, 430 proteins were identified, including 34 that were undetectable in untreated serum samples. This validated the high sensitivity and specificity of this method in serum proteomics, demonstrating its potential for isolating and identifying low-abundance proteins in complex biological samples. The developed system offers a valuable tool for advancing human serum proteomics research and targeted protein analysis.

CAS / MDL: 9002-98-6
研究领域: 生物化学 · 纳米技术 · 蛋白质组学 · 生物医学工程 · 分子生物学
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Journal of Immunotoxicology · 2019 · IF 1.734

Improvement and optimization of a T-cell-dependent antibody response (TDAR) method for BALB/c mice using keyhole limpet hemocyanin (KLH) as specific antigen

Penghuan Chang,Ling Huang,Mianqing Huang,Shuhong Tian,Zhaoxin Yang

Although T-cell-dependent antibody response (TDAR) assays with keyhole limpet hemocyanin (KLH) as specific antigen have many advantages, most experiments produce qualitative results based on antibody titers. It was hypothesized that if experimental conditions (like antigen coating concentration, serum dilution, and detecting [here, horseradish peroxidase-goat anti-mouse IgG] antibody dilution) could be optimized, the resulting measured value (here, optical density) could be used to directly analyze and evaluate the experimental results. This means specifically that the assay OD values could be used for approximate quantitative statistical analysis; it does not require a further conversion of the data into qualitative forms or require obtaining further titer data from additional experiments. As such, the use of this “improved” assay would: greatly reduce the complexity of experimental operations; improve overall sensitivity and practicality of traditional TDAR assays; and, allow for direct assessing of any immunosuppression caused by a test drug in a host. Here, KLH-immunized serum was obtained from BALB/c mice, and the means to detect serum anti-KLH antibodies by an indirect ELISA were optimized. The results indicated that in this system, the optimal KLH coating concentration was 80 μg/ml, the optimal dilution range of the serum (at immunization dose of 5 mg KLH/kg) was 1:200–1:800, and the optimal dilution of HRP-goat anti-mouse IgG antibody was 1:16,000. Methodology verification was performed and a regression model of y = 144.16x + 0.9815 (R2 = 0.9571, indicating very good linearity) was obtained. Intragroup precision was 7.51–9.40%; the intergroup coefficient of variation was 9.83–14.22%. The lower limit of detection was 0.1385. The present results showed this indirect ELISA exhibited very good linearity, accuracy, and precision.

CAS / MDL: 9005-64-5
研究领域: 免疫学 · 生物化学 · 药理学
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FOOD RESEARCH INTERNATIONAL · 2025 · IF 8

Ultrasonic regulation of β-lactoglobulin/puerarin complex: structural modification, digestive properties, and allergenicity suppression through controlled polyphenol interactions

Peng Man,Jinghua Pei,Sijia Chen,Xiue Han,Huajiang Zhang

This study explored the effects of ultrasound on β-lactoglobulin (β-LG)/puerarin (PUE) complexes, focusing on structural, digestive, and allergenic properties. Molecular simulations revealed hydrogen bonding as the primary interaction between PUE and β-LG. Ultrasound reduced β-LG crystallinity (via XRD), further enhanced by PUE encapsulation. Moderate ultrasound (300 W) increased thermal denaturation temperature by 8.2 °C (DSC) and enhanced digestibility to 78.3 ± 1.2 % (vs. 62.5 ± 2.1 % in untreated group), while excessive power (500 W) caused structural damage. Ultrasonic treatment increased the β-sheet content of β-LG, led to a reorganization of its tertiary structure. Allergenicity assays demonstrated significant reductions in IgE and IgG binding capacity for 300 W-treated complexes. These findings highlighted moderate ultrasound (300 W) as an effective strategy to modulate functionality and reduce allergenicity in protein-polyphenol colloids, offered insights for food science applications.

CAS / MDL: 7647-14-5
研究领域: 食品科学 · 超声学 · 蛋白质化学 · 生物化学 · 致敏性研究 · 分子模拟 · 消化生理学
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FOOD CHEMISTRY · 2025 · IF 9.8

Investigation of α-dicarbonyl compounds induced glycation on the digestive stability of shrimp tropomyosin and intestinal microbiota

Lichun Liu,Ziye Zhang,Wei Yang,Fatih Özogul,Zhenxing Li,Hong Lin

α-dicarbonyl compounds are widely present in foods, while their modification effects on food allergens are unclear. This work was conducted to investigate the effects of α-dicarbonyl compounds (glyoxal (GO), methylglyoxal (MGO) and 2, 3-butanedione (BU)) on the digestive stability and intestinal flora of shrimp tropomyosin (TM). As the results, higher α-dicarbonyl concentration, longer time and higher temperature could exacerbate the modification of TM. Compared with TM, GO and MGO significantly reduced the digestive resistance of TM and the IgG-binding capacity of AGEs-TM digests. Although in vitro digestion reduced IgE-binding capacity of TM/AGEs-TM, in vivo evidence (intestinal biopsy and gut microbiota) suggested that they still elicits immune responses. In addition, AGEs-TM could reduce the richness and diversity of intestinal flora, increase the abundance of inflammation-related flora and flora imbalance index, which could provide a new insight into the effects of AGEs on the digestive stability of shrimp tropomyosin and intestinal microbiota.

研究领域: 食品化学 · 过敏学 · 分子生物学 · 营养科学 · 微生物学
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BIOSENSORS & BIOELECTRONICS · 2024 · IF 10.7

Antibody screening-assisted multichannel nanoplasmonic sensing chip based on SERS for viral screening and variants identification

Yi Liu,Huanjiao Weng,Zhiwei Chen,Ming Zong,Shubin Fang,Zili Wang,Shaohua He,Yangming Wu,Jizhen Lin,Shangyuan Feng,Duo Lin

The Omicron variants of SARS-CoV-2 have been spreading globally and has never disappeared from our sight, indicating that its coexistence with humans has become a fact, and monitoring its evolution and spread remains a current task. Although polymerase chain reaction (PCR) is the most commonly used virus detection method, it requires labor-intensive and time-consuming procedures in a laboratory setting. Herein, a multichannel nanoplasmonic sensing chip based on surface enhanced Raman spectroscopy (SERS) was developed for detecting N and S proteins, as well as IgG and IgM, related to SARS-CoV-2 Omicron variants. Through a self-screening process, specific antibodies for on-site and rapid identification of important variants of concerns (VoCs) were obtained, and their binding was confirmed by protein structure analysis. The use of these S protein specific antibodies can accurately identify Omicron VoCs (BA. 5, BF.7,XBB.1.5) with the detection limit (LoD) at 0.16 pg/mL. Then, the proposed SERS array chip was integrated with a hand-held Raman spectrometer to successfully detect the Omicron subvariants in real saliva samples within only 20 minutes, greatly reducing the detection time of PCR. This sensing technology will provide a powerful and rapid point-of-care testing (POCT) method for virus diagnosis, subtype identification, and post infection antibody level monitoring.

研究领域: 生物医学工程 · 分析化学 · 病毒学 · 医学诊断
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SENSORS AND ACTUATORS B-CHEMICAL · 2020 · IF 7.1

Ultrahigh-sensitivity label-free optical fiber biosensor based on a tapered singlemode- no core-singlemode coupler for Staphylococcus aureus detection

Ling Chen,Yuan-Kui Leng,Bin Liu,Juan Liu,Sheng-Peng Wan,Tao Wu,Jinhui Yuan,Liyang Shao,Guoqiang Gu,Yong Qing Fu,Hengyi Xu,Yonghua Xiong,Xing-Dao He,Qiang Wu

An ultra-high sensitivity label-free optical fiber biosensor for inactivated Staphylococcus aureus ( S. aureus ) detection is proposed and investigated in this study, with additional advantages of robust and stability compared to traditional tapered fiber structure. The proposed fiber biosensor is based on a tapered singlemode- no core-singlemode fiber coupler (SNSFC) structure, where the no core fiber was tapered to small diameter (taper-waist diameter of about 10 μm) and functionalized with the pig immunoglobulin G (IgG) antibody for detection of S. aureus . The measured maximum wavelength shift of the sensor for an S. aureus concentration of 7 × 10 1 CFU/mL (colony forming unit per milliliter) is 2.04 nm, which is equivalent to a limit of detection (LOD) of 3.1 CFU/mL (a highest LOD reported so far for optical fiber biosensors), considering the maximum wavelength variation of the sensor in phosphate buffered saline (PBS) is ±0.03 nm over 40 min, where 3 times of maximum wavelength variation (3 × 0.03 = 0.09 nm) is defined as measurement limit. The response time of the developed fiber sensor is less than 30 min. The ultra-sensitive biosensor has potential to be widely applied to various areas such as disease, medical diagnostics and food safety inspection.

研究领域: 生物传感 · 光纤技术 · 微生物学 · 食品安全 · 医学诊断
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TALANTA · 2017 · IF 4.162

A sensitive SPR biosensor based on hollow gold nanospheres and improved sandwich assay with PDA-Ag@Fe3O4/rGO

Shuo Li,Qiong Wu,Pinyi Ma,Yue Zhang,Daqian Song,Xinghua Wang,Ying Sun

A novel surface plasmon resonance (SPR) biosensor based on hollow gold nanospheres (HGNPs) and an improved sandwich assay was developed to detect rabbit IgG. The electromagnetic coupling between the HGNPs and Au film, and the notable plasmonic fields spread over the inner and outer surfaces of HGNPs, led to the considerable amplification of the SPR signal. [email protected] 3 O 4 /reduced graphene oxide ( [email protected] 3 O 4 /rGO) was introduced to bind detection antibody (Ab 2 ) to form the improved sandwich structure. Ag nanoparticles were excited to produce SPR and their hot electrons were doped on graphene thin films, which amplified the response of biomolecules. Magnetic nanoparticles (Fe 3 O 4 ) simplified the collection of Ab 2 [email protected] 3 O 4 /rGO. An external layer of polydopamine (PDA) permitted the efficient immobilization of Ab 2 without activation via abundant functional groups and protected the nanoparticles from etching or agglomeration. In addition, because of its large mass, Ab 2 [email protected] 3 O 4 /rGO also played a key role in the further amplification of the SPR response signals. This novel SPR biosensor exhibited an effective response to the rabbit IgG at the different concentrations ranging from 0.019 to 40.00   μg   mL −1 . This value is 132 times lower than that observed for a traditional SPR biosensor based on Au-3-mercaptopropionic acid and 8 times lower than that obtained from an Ab 2 sandwich assay, which indicates that the SPR sensor has high sensitivity. In addition, the designed biosensor showed satisfactory recoveries to detect the rabbit IgG spiked in serum samples. Therefore, the novel SPR biosensor with high sensitivity and acceptable recovery has potential for practical applications.

研究领域: 纳米技术 · 生物传感器 · 分析化学 · 材料科学 · 生物医学工程
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES · 2024 · IF 7.7

The synthesis and preliminary immunological evaluation of a dual-adjuvant SARS-CoV-2 RBD vaccine: Covalent integration of TLR7/8 and iNKT cell agonists

Ying Xu,Jing Yang,Xiongyan Meng,Shuai Meng,Tiantian Sun,Ning Ding,Chengli Zong

Covalently linking an adjuvant to an antigenic protein enhances its immunogenicity by ensuring a synergistic delivery to the immune system, fostering a more robust and targeted immune response. Most adjuvant-protein conjugate vaccines incorporate only one adjuvant due to the difficulties in its synthesis. However, there is a growing interest in developing vaccines with multiple adjuvants designed to elicit a more robust and targeted immune response by engaging different aspects of the immune system for complex diseases where traditional vaccines fall short. Here, we pioneer the synthesis of a dual-adjuvants protein conjugate Vaccine 1 by assembling a toll-like receptor 7/8 (TLR7/8) agonist, an invariant natural killer T cell (iNKT) agonist with a clickable bicyclononyne (BCN). The BCN group can bio-orthogonally react with azide-modified severe acute respiratory syndrome coronavirus-2 receptor-binding domain (SARS-CoV-2 RBD) trimer antigen to give the three-component Vaccine 1 . Notably, with a mere 3 μg antigen, it elicited a balanced subclass of IgG titers and 20-fold more IgG2a than control vaccines, highlighting its potential for enhancing antibody-dependent cellular cytotoxicity. This strategy provides a practicable way to synthesize covalently linked dual immunostimulants. It expands the fully synthetic self-adjuvant protein vaccine that uses a single adjuvant to include two different types of adjuvants.

研究领域: 免疫学 · 病毒学 · 药物化学 · 疫苗研发
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