CHEMBIOCHEM
·
2024
·
IF 2.6
Xiang Zhu,Weizhe Luo,Donghui Zhang,Runhui Liu
With the increasing use of polyethylene glycol (PEG) based proteins and drug delivery systems, anti-PEG antibodies have commonly been detected among the population, causing the accelerated blood clearance and hypersensitivity reactions, poses potential risks to the clinical efficacy and safety of PEGylated drugs. Therefore, vigilant monitoring of anti-PEG antibodies is crucial for both research and clinical guidance regarding PEGylated drugs. The enzyme-linked immunosorbent assay (ELISA) is a common method for detecting anti-PEG antibodies. However, diverse coating methods, blocking solutions and washing solutions have been employed across different studies, and unsuitable use of Tween 20 as the surfactant even caused biased results. In this study, we established the optimal substrate coating conditions, and investigated the influence of various surfactants and blocking solutions on the detection accuracy. The findings revealed that incorporating 1% bovine serum albumin into the serum dilution in the absence of surfactants will result the credible outcomes of anti-PEG antibody detection.
研究领域:
药学
·
免疫学
·
药物递送系统
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DOI
PubMed
高影响因子
MOLECULAR PHARMACEUTICS
·
2024
·
IF 4.5
Qiuning Zhang,Lei Li,Qianqian Yang,Wei Chen,Ziyuan Wang,Mingcui Zhang
The accurate and efficient quantification of nanodrug dosage is crucial for early anticancer therapy. The enzyme-linked immunosorbent assay (ELISA) has emerged as a robust tool for detecting anticancer nanodrug dosage; however, the development of sensing elements to quantify anticancer nanodrugs still poses a challenge. To overcome this problem, we utilize polysuccinimide-loaded curcumin (CUR @PSIOAm) as a model to employ an ELISA based on peroxidase nanozyme Pt-SiO2 Janus nanoparticles (Pt-SiO2 JNPs) for the indirect quantitative analysis of intracellular anticancer nanodrug dosage. This novel approach employs an immunoassay to indirectly quantify the dosage of anticancer nanodrugs while preserving its structural integrity. The silica components of Pt-SiO2 JNPs adsorb intermediates, while the Pt NP components exhibit high catalytic activity. Pt-SiO2 JNPs are functionalized with anti-PSIOAm antibody (Pt-SiO2 JNPs-Ab) to serve as an immunosensor capable of specific recognition of CUR @PSIOAm. Additionally, we employed cytotoxicity assays and confocal imaging techniques to demonstrate the excellent biocompatibility of CUR @PSIOAm, as well as its specific uptake by cancer cells. According to the experimental results, the limit of detection (LOD) for the immunoassay of Pt-SiO2 JNPs as a marker for detecting CUR @PSIOAm is approximately 4.5-fold lower than that of horseradish peroxidase. Therefore, by optimizing the conditions, we established a direct competitive ELISA using Pt-SiO2 JNPs as colorimetric indicators for the quantitative detection of intracellular CUR @PSIOAm. The LOD for this ELISA was determined to be 0.01 ng/mL, while the loaded CUR amount calculated from the drug loading capacity was found to be 0.22 pg/mL. Furthermore, the recoveries obtained from this established ELISA ranged between 94.0 and 108%, demonstrating excellent accuracy. Consequently, the peroxidase mimic Pt-SiO2 JNPs-based ELISA exhibits significant potential for precise quantification of intracellular anticancer nanodrug dosages.
研究领域:
生物医学工程
·
肿瘤学
·
分析化学
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DOI
PubMed
高影响因子
Analytical Methods
·
2025
·
IF 2.6
Jiexin Zheng,Tianyu Lan,Bo Xu,Chunlei Cao,Feng Sun,Yue He,Qiyang Zhao,Jing Li,Yaohai Zhang,Chengqiu Wang,Yongliang Cui
Obacunone, a limonoid compound structurally related to limonin, is commonly found in plants of the Rutaceae family, including Phellodendron and Citrus species. Obacunone serves as a marker for assessing the quality of herbal medicines and traditional Chinese medicine (TCM) formulations. However, the current detection method for obacunone requires expensive equipment, such as high-performance liquid chromatography (HPLC) and ultra-performance liquid chromatography-mass spectrometry (UPLC-MS), and lacks convenient and rapid detection alternatives. In this study, we developed an indirect competitive enzyme-linked immunosorbent assay (icELISA) for obacunone detection. The half-maximal inhibitory concentration (IC50) for the icELISA was established at 0.44 ng/mL, with a detection range extending from 0.09 ng/mL to 3.52 ng/mL. The spike-recovery experiments conducted demonstrated excellent method stability and reliability, with recovery rates consistently ranging from 94.6% to 105.5%, well within the acceptable criteria for analytical method validation in pharmaceutical analysis. The obacunone content in seven TCM formulations and two herbal medicine samples was analyzed using both icELISA and UPLC-MS, demonstrating consistent results between the two analytical methods. The results demonstrated that the developed immunoassay method exhibited excellent applicability for obacunone detection in both herbal medicines and TCM formulations, with significant advantages in terms of analytical efficiency and stability.
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产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2016
·
IF 4.712
Yi Liang,Xiaolin Huang,Ruijin Yu,Yaofeng Zhou,Yonghua Xiong
The present study described a novel fluorescence enzyme-linked immunosorbent assay (ELISA) used to detect ochratoxin A (OTA) by using the glucose oxidase (GOx)-mediated fluorescence quenching of mercaptopropionic acid-capped CdTe quantum dots (MPA-QDs), in which GOx was used as an alternative to horseradish peroxidase (HRP) for the oxidization of glucose into hydrogen peroxide (H 2 O 2 ) and gluconic acid . The MPA-QDs were used as a fluorescent signal output, whose fluorescence variation was extremely sensitive to the presence of H 2 O 2 or hydrogen ions in the solution. Under the optimized conditions, the proposed fluorescence ELISA demonstrated a good linear detection of OTA in corn extract from 2.4 pg mL −1 to 625 pg mL −1 with a limit of detection of 2.2 pg mL −1 , which was approximately 15-fold lower than that of conventional HRP-based ELISA. Our developed fluorescence immunoassay was also similar to HRP-based ELISA in terms of selectivity , accuracy, and reproducibility. In summary, this study was the first to use the GOx-mediated fluorescence quenching of QDs in immunoassay to detect OTA, offering a new possibility for the analysis of other mycotoxins and biomolecules.
研究领域:
食品科学
·
分析化学
·
毒理学
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DOI
高影响因子
Journal of Food Measurement and Characterization
·
2024
·Vol. 18
·期 4
·2561-2572
·
IF 2.9
Liu Junwei,Wu Yuqing,Liu Chenchen,Cai Yan,Zhang Rui,Lu Yang
Based on the new hapten of fipronil, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and time-resolved fluorescent immunochromatographic assay (TRFICA) for the determination of fipronil were developed. The sensitivity (IC 50 ) of the ic-ELISA method was 2.33 ± 0.14 ng mL −1 (n = 3), and the limit of detection (LOD) (IC 20 ) was 0.27 ± 0.02 ng mL −1 (n = 3). The cross-reaction rates of the three fipronil metabolites were all higher than 66.7%, and those of the three analogs were no more than 0.05%. The TRFICA was optimized with a handheld reader. The LOD was 0.862 ng mL −1 (n = 3). In this study, the pretreatment methods of samples such as Chinese cabbage, spinach, apple and egg, were explored. Methanol and acetonitrile were used to extract different food samples and the diluents were optimized. Three concentration levels in the spiked food samples were detected by the two methods. The range of recovery was 88.65–116.32% for ic-ELISA and 95.48–112.61% for TRFICA. In this study, the accuracy of the two methods was also judged by testing commercial quality control samples. The results showed that the accuracy was 83.75%–113.33% for ic-ELISA and 98.33–107.13% for TRFICA.
研究领域:
分析化学
·
食品安全检测
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DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Yarong Ren,Juan Wei,Ye Wang,Peng Wang,Yanwei Ji,Baoyuan Liu,Jianlong Wang,Gualberto González-Sapienza,Yanru Wang
Salmonella are major pathogens that cause foodborne diseases. In this work, a broad-spectrum Salmonella nanobody-01 (Nb-01) was isolated and applied in the development of a streptavidin-bridged sandwich ELISA (SAB-ELISA) for simultaneously identifying five Salmonella serovars , including Salmonella Enteritidis ( S. Enteritidis), Salmonella Typhimurium ( S. Typhimurium), Salmonella London (S. London), Salmonella Paratyphi B (S. Paratyphi B) and Salmonella Hadar (S. Hadar). In this work, streptavidin (SA) was utilized as a scaffold to directionally immobilize biotinylated nanobody (BiNb) and Salmonella was detected by phage-displayed nanobodies. The SAB-ELISA can be accomplished within 180 min with a limit of detection (LOD) of 6.31 × 10 3 colony forming units (CFU) mL −1 , 9.15 × 10 3 CFU mL −1 , 4.23 × 10 3 CFU mL −1 , 7.31 × 10 3 CFU mL −1 and 7.25 × 10 3 CFU mL −1 towards S. Typhimurium, S. Enteritidis, S. London, S. Paratyphi B and S. Hadar, respectively. In comparison of sandwich ELISA by passive immobilization of Nb-01 on polystyrene plate, the sensitivity was increased by around 6-fold, which confirmed the enhanced immobilization efficacy of SAB-ELISA. Furthermore, the feasibility of the assay for S. Typhimurium determination in actual samples was evaluated, showing excellent recovery, inter-day and intra-day precision.
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2021
·
IF 6.986
Hailing Zheng,Hailiang Yang,Yang Zhou,Tianxiao Li,Qinglin Ma,Bing Wang,Qin Fang,Haixiang Chen
The extraction and identification of silk residues in tombs is of great significance for studying the distribution and spread of early silk. However, the complex organic matter in the tomb hinders the accurate identification of silk. In this study, a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) based on immunomagnetic beads (IMBs) was developed for the rapid enrichment and detection of silk residues. The double-antibody sandwich ELISA method established by pairing the IMBs prepared by the silk fibroin monoclonal antibody SF-3 and the silk fibroin monoclonal-labeled antibody bio-SF-1 had the highest detection sensitivity, with a linear detection range of 10 to 104 ng mL–1 and a detection limit of 5.12 ng mL–1. This method was excellent in the extraction and analysis of silk residues from archaeological imprints and soil samples and successfully identified silk residues in samples at the final stage of silk degradation (physical invisible silk). The proteomics analysis results demonstrated the feasibility and practicability of this method.
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2021
·Vol. 188
·期 8
·1-11
·
IF 5.833
Feng Jun,Lu Hao,Yang Yu,Huang Wenyi,Cheng Hao,Kong Hongxing,Li
Lijun
By in situ synthesis of gold nanoparticles (AuNPs) within the acid-etched (AE) MIL-101 (Cr) framework, AE-MIL-101 (Cr) nanocomposites embedded with AuNPs (AuNP/AE-MIL-101 (Cr)) were prepared as surface-enhanced Raman scattering (SERS) substrate. AuNPs are uniformly distributed and stabilized inside the metal-organic framework (MOF), thus forming more SERS hotspots. The SERS performance of AuNP/AE-MIL-101 (Cr) was evaluated using 4-mercaptophenylboronic acid (4-MPBA), 4-mercaptobenzoic acid (4-MBA), benzidine, and rhodamine 6G (R6G). The SERS substrate displays satisfying stability with very low background signal. When benzidine is used as the Raman reporter, the limit of detection (LOD) can reach 6.7 × 10 −13 mol·L −1 , and the relative standard deviation (RSD) of the intra- and inter-batch repetitive tests is less than 5.2%. On this basis, we developed a method for the detection of human carboxylesterase 1 (hCE 1) in human serum using AuNP/AE-MIL-101 (Cr) nanocomposite as SERS substrate and enzyme-linked immunosorbent assay (ELISA) colorimetric substrate as SERS marker. This method was used to determine hCE 1 in clinical serum samples without complicated sample pretreatment, and the detection results were consistent with the data determined by ELISA. In the concentration range 0.1–120 ng·mL −1 , the SERS signal intensity of benzidine at 1609 cm −1 gradually decreases with the increase of hCE 1 concentration (R 2 = 0.9948). The average recoveries of hCE 1 in human serum are in the range 84 to 108%, with RSDs lower than 7.7%. Graphical abstract
By using AuNP/acid etching-MIL-101(Cr) metal organic framework (MOF) as SERS substrate and enzyme-linked immunosorbent assay (ELISA) colorimetric substrate as the SERS marker, a rapid and sensitive method for the determination of human carboxylesterase 1 (hCE1) in human serum samples has been developed.
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DOI
PubMed
高影响因子
Food Chemistry-X
·
2024
·
IF 6.5
Lei Zhang,Huiqiang Huang,Mengyao Liao,Chenghua Luo,Qiuyue Wu,Rentang Huang,Hongmei Liu,Xiangsheng Zhao,Shumei Wang
The enzyme-linked immunosorbent assay (ELISA) is a powerful and high-throughput method for detecting aflatoxin B 1 (AFB 1 ) in food. However, the susceptibility of native enzymes to environmental factors limits their potential applications. This study introduces the combined use of precious metal nanozymes as alternative catalysts to improve the performance of traditional ELISA. Au@Pt can be linked to antibodies to construct recognition probes, onto which platinum nanoparticles (PtNPs) are ingeniously deposited in situ to amplify the detection signal, thereby increasing the sensitivity. The proposed PtNP-enhanced Au@Pt-ELISA exhibited excellent resistance against matrix interference for AFB 1 detection in lotus seed powder and achieved a detection limit of 1.6 pg/mL (0.32 μg/kg), which was approximately six times lower than that of traditional ELISA. Furthermore, the results of the proposed approach were strongly correlated with LC–MS/MS analysis results for real samples ( r = 0.9888, n = 13). This study offers a new strategy for AFB 1 monitoring in a complex matrix.
研究领域:
食品科学
·
免疫学
·
纳米技术
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产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2021
·Vol. 13
·期 14
·1757-1765
·
IF 2.896
Can Zhang,Xiaoxiao Wu,Dongyang Li,Jinnuo Hu,Debin Wan,Zhen Zhang,Bruce D. Hammock
As a major metabolite of pyrethroid pesticides, 3-phenoxybenzoic acid (3-PBA) can be an indicator of health risk and human exposure assessment. Based on nanobodies (Nbs), we have developed a rapid flow-through dot enzyme linked immunosorbent assay (dot ELISA) and gold nanoparticle (GNP) lateral-flow immunoassay for detecting 3-PBA. The limit of detection (LOD) values for detecting 3-PBA by flow-through dot ELISA and GNP lateral-flow immunoassay were 0.01 ng mL−1 and 0.1 ng mL−1, respectively. The samples (urine and lake water) with and without 3-PBA were detected by both nanobody-based flow-through dot ELISA and GNP lateral-flow immunoassay, as well as liquid chromatography-mass spectrometry (LC-MS) for validation. There was good consistency between the results of the immunoassays. This demonstrated that the two developed nanobody-based immunoassays are suitable for rapid detection of 3-PBA.
研究领域:
环境科学
·
分析化学
·
免疫学
·
纳米技术
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DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2026
·
IF 9.8
Boyan Sun,Linqian Qin,Vasily G. Panferov,Chujun Liu,Xiaomei Wang,Jiankang Ren,Qian Ran,Sihan Wang,Haiyang Jiang
Ractopamine (RAC), a banned β-adrenergic agonist, poses severe risks to human health, necessitating rapid and ultrasensitive detection methods. This work presents a novel dual-mode immunoassay utilizing peroxidase-mimicking Au@Pt core-shell nanozymes for the colorimetric and photothermal detection of RAC. The optimized one-step synthesized Au@Pt nanoparticles (Au/Pt = 1:1) exhibited exceptional peroxidase-like activity (27.32 U/mg). After conjugation with monoclonal antibodies, the nanozymes served as immunoprobes in a one-step enzyme-linked immunosorbent assay (ELISA). The enhanced catalysis was elucidated by reactive oxygen species trapping, confirming hydroxyl radicals and singlet oxygen as key intermediates. The assay demonstrated outstanding sensitivity, with limits of detection at 5.32 pg/mL (colorimetric) and 1.03 pg/mL (photothermal), a significant improvement over conventional ELISA. Practical applicability was confirmed in spiked pork and pig urine, achieving recoveries of 86.0–113.0% (coefficient of variation<14.3%), offering a robust platform for on-site screening.
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产品关联
DOI
PubMed
高影响因子
Food Bioscience
·
2026
·
IF 6.2
Shuai Zhao,Yunfei Xia,Xing Chen,Yan Chen,Xuezhuang He,Menqing Cui,Yuchen Gao,Lei Wang,Jing Lu,Yawei Shi
This study aimed to elucidate the protein profiles of low-temperature Daqu during the Daqu-making process. Temporal metaproteomics was conducted across 8 stages of Daqu production. The top 50 proteins in Daqu were primarily derived from peas, the diversity and abundance of microbial-source proteins increased significantly from the Liangmei stage. The marker microbial proteins at different Daqu-making stages were screened via random forest analysis. Chaperonin GroEL from Lactiplantibacillus plantarum and Enolase from Lactobacillus acidophilus were identified as markers during Liangmei and Chaohuo stages. Additionally, the enzyme activities of hydrolases and oxidoreductases were analyzed, suggesting that the activities of acid protease, polygalacturonase, and carboxyl esterase significantly improved after the Liangmei stage. Notably, ELISA (enzyme-linked immunosorbent assay) for superoxide dismutase and α-galactosidase was first applied to the biochemical analysis of low-temperature Daqu, and verified its reliability and applicability. This study was conducive to understanding the protein dynamics during Daqu-making, and provided insights for evaluating the quality of Daqu.
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高影响因子
危险品化学品经营许可证（带存储）