FOOD AND AGRICULTURAL IMMUNOLOGY
·
2018
·
IF 2.568
Lu Zeng,Wei Jiang,Liqiang Liu,Shanshan Song,Hua Kuang
Vitamin B2 (riboflavin) is a water-soluble vitamin that has important roles in human health. In this study, we developed a sensitive and specific monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and lateral-flow immunochromatographic assay (ICA) strip for the rapid detection of vitamin B2. Following routine fusion and selection, the optimum monoclonal antibody against vitamin B2 was obtained. The 50% inhibitory concentration and limit of detection of ic-ELISA were 8.18 and 1.80 ng/mL, respectively. The cut-off value of the lateral-flow ICA strip was 50 ng/mL. The results revealed that our developed methods are suitable for the on-site detection and mass screening of vitamin B2 in food and pharmaceutical products.
研究领域:
分析化学
·
免疫学
·
食品科学
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产品关联
DOI
高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
·
2020
·
IF 4.872
Zi-Jian Chen,Xi-Xia Liu,Zhi-Li Xiao,Hui-Jun Fu,Yu-Ping Huang,Shu-Yi Huang,Yu-Dong Shen,Fan He,Xing-Xing Yang,Bruce Hammock,Zhen-Lin Xu
1-naphthol (1-NAP) is the main metabolite of pesticide carbaryl and naphthalene , and is also a genotoxic and carcinogenic intermediate in the synthesis of organic compound, dyes, pigment and pharmaceutical industry . In this work, two novel haptens were designed and synthesized for developing a competitive indirect enzyme-linked immunosorbent assay (ciELISA) method for 1-NAP in urine samples. The assay showed a limit of detection of 2.21 ng/mL and working range from 4.02 ng/mL to 31.25 ng/mL for 1-NAP in optimized working buffer. The matrix effect of samples was eliminated via 15-fold dilution of optimized working buffer. Good average recoveries (102.4%–123.4%) with a coefficient of variation from 11.7% to 14.7% was obtained for spiked urine samples. Subsequent instrument verification test showed good correlation between the results of ciELISA and high-performance liquid chromatography . The developed ciELISA is a high-throughput tool to monitor 1-NAP in urine, which can provide technical support for the establishment of biological exposure level for the exposure to carbaryl, naphthalene and other related pollutants.
研究领域:
环境生物学
·
毒理学
·
分析化学
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DOI
PubMed
高影响因子
Foods
·
2022
·Vol. 11
·期 3
·335
·
IF 5.561
Disha Lu,Xu Wang,Ruijue Su,Yongjian Cheng,Hong Wang,Lin Luo,Zhili Xiao
A novel and efficient immunoaffinity column (IAC) based on bispecific monoclonal antibody (BsMAb) recognizing aflatoxin B1(AFB1) and ochratoxin A (OTA) was prepared and applied in simultaneous extraction of AFB1and OTA from food samples and detection of AFB1/OTA combined with ic-ELISA (indirect competitive ELISA). Two deficient cell lines, hypoxanthine guanine phosphoribosyl-transferase (HGPRT) deficient anti-AFB1hybridoma cell line and thymidine kinase (TK) deficient anti-OTA hybridoma cell line, were fused to generate a hybrid-hybridoma producing BsMAb against AFB1and OTA. The subtype of the BsMAb was IgG1via mouse antibody isotyping kit test. The purity and molecular weight of BsMAb were confirmed by SDS-PAGE method. The cross-reaction rate with AFB2was 37%, with AFG115%, with AFM148%, with AFM210%, and with OTB 36%. Negligible cross-reaction was observed with other tested compounds. The affinity constant (Ka) was determined by ELISA. The Ka (AFB1) and Ka (OTA) was 2.43 × 108L/mol and 1.57 × 108L/mol, respectively. Then the anti-AFB1/OTA BsMAb was coupled with CNBr-Sepharose, and an AFB1/OTA IAC was prepared. The coupling time and elution conditions of IAC were optimized. The coupling time was 1 h with 90% coupling rate, the eluent was methanol–water (60:40,v:v, pH 2.3) containing 1 mol/L NaCl, and the eluent volume was 4 mL. The column capacities of AFB1and OTA were 165.0 ng and 171.3 ng, respectively. After seven times of repeated use, the preservation rates of column capacity for AFB1and OTA were 69.3% and 68.0%, respectively. The ic-ELISA for AFB1and OTA were applied combined with IAC. The IC50(50% inhibiting concentration) of AFB1was 0.027 ng/mL, the limit of detection (LOD) was 0.004 ng/mL (0.032 µg/kg), and the linear range was 0.006 ng/mL~0.119 ng/mL. The IC50of OTA was 0.878 ng/mL, the LOD was 0.126 ng/mL (1.008 µg/kg), and the linear range was 0.259 ng/mL~6.178 ng/mL. Under optimum conditions, corn and wheat samples were pretreated with AFB1-OTA IAC. The recovery rates of AFB1and OTA were 95.4%~105.0% with ic-ELISA, and the correlations between the detection results and LC-MS were above 0.9. The developed IAC combined with ic-ELISA is reliable and could be applied to the detection of AFB1and OTA in grains.Keywords:aflatoxin B1;ochratoxin A;bispecific monoclonal antibody;immunoaffinity column;ic-ELISA
研究领域:
食品科学
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分析化学
·
毒理学
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DOI
PubMed
高影响因子
Applied Sciences-Basel
·
2018
·Vol. 8
·期 4
·560
·
IF 1.689
Guangyang Liu,Yongxin She,Sihui Hong,Jing Wang,Donghui Xu
Featured ApplicationThe developed method would be useful for monitoring melamine in milk and formula powdered milk.AbstractWe present a directly competitive fluorescence assay for highly sensitive detection of melamine in milk using magnetic dummy molecularly imprinted polymers (MDMIPs). The detection principle is based on competitive binding between the fluorescent label and melamine on the MDMIPs. The fluorescent label was obtained by combining fluorescein isothiocyanate (FITC) with melamine in ethanol and water. MDMIPs were prepared on the surface of Fe3O4@SiO2nanoparticles using 2,4-diamino-6-methyl-1,3,5-triazine as dummy template. The MDMIPs were characterized and their adsorption capacity was evaluated based on their static adsorption and Scatchard analysis. Results suggest that MDMIPs were successfully coated on the Fe3O4@SiO2surface and had a core–shell structure. Adsorption experiments suggested that the MDMIPs had higher specific recognition capacities for melamine and FITC–melamine (FITC-Mel) than did magnetic dummy molecularly non-imprinted polymers. Competitive binding between FITC-Mel and melamine was performed under the optimum conditions to determine melamine quantitatively. The linear range of this fluorescence assay was 0.1–20 mg/L for melamine detection. The detection limit was 0.05 mg/L in negative milk samples. The assay was also successfully employed to detect melamine in spiked milk samples, with satisfactory recoveries, i.e., between 70.2% and 92.7%.Keywords:ELISA-like fluorescence assay;magnetic dummy molecularly imprinted polymers;2,4-diamino-6-methyl-1,3,5-triazine;fluorescein isothiocyanate;melamine
研究领域:
分析化学
·
食品安全检测
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产品关联
DOI
高影响因子
TALANTA
·
2022
·
IF 6.556
Xin Wang,Min Zhang,Xiaoxu Pang,Kunlun Huang,Zhiyi Yao,Xiaohong Mei,Nan Cheng
Staphylococcus aureus is a frequent pathogenic bacterium that has a significant detrimental influence on the health of the human body. Therefore, developing a practical and portable detection platform is critical to ensuring food safety. Nanozymes are a kind of engineered nanomaterials with superior enzyme-like activities, providing infinite possibilities for the development of highly sensitive analytical assays. In this study, mesoporous core−shell [email protected] ( [email protected] ) nanozymes were synthesized and then applied as a signal amplifier in Staphylococcus aureus colorimetric immunoassay . At the same time, a careful comparative study of the catalytic performance with natural horseradish peroxidase (HRP), [email protected] nanozymes and their complexes [email protected] (HRP coupling with [email protected] nanozymes) were firstly performed, as well as clever using a colorimeter to achieve portable signal output. [email protected] bioprobes enable remarkable peroxidase-like catalytic activity, resulting in the highest sensitivity with a limit of detection (LOD) improved from 1 × 10 5 to 1 × 10 3 CFU/mL. In all, this work conducted a valuable comparative study and presented an improved strategy for the portable detection of pathogenic bacteria, which was expected to be used in a wide range of applications in food safety regulation and biomedicals arenas.
研究领域:
分析化学
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生物技术
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食品安全
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生物传感器
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纳米技术
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产品关联
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2022
·
IF 5.304
Hao Lu,Yu Yang,Ruijue Chen,Wenyi Huang,Hao Cheng,Xueping Liu,Hongxing Kong,Lijun Li,Jun Feng
In this study, we synthesized gold nanoparticles (AuNPs) in situ on the surface of two-dimensional Cu-TCPP (meso-tetra(4-carboxyphenyl)porphine) metal– organic framework (MOF) nanosheets to produce AuNP@Cu-TCPP nanocomposites as surface-enhanced Raman scattering (SERS) substrates. The SERS signal was enhanced in two ways: aggregation of the AuNPs@Cu-TCPP near the SERS hotspot through the strong adsorption capacity of the MOF, and the synergistic effect between the charge-transfer enhancement of the Cu-TCPP MOF and the electromagnetic enhancement of plasmonic metal AuNPs. The SERS activity of AuNPs@Cu-TCPP was evaluated by the analysis of multiple model analytes; Rhodamine 6G (R6G) and benzidine were detected with detection limits below 10 –13 and 10 –12 mol·L –1 , respectively. We used AuNP@Cu-TCPP as a SERS substrate combined with an enzyme-linked immuno- sorbent assay (ELISA) to detect the tumor marker carboxylesterase 1 (hCE1) in human serum. The SERS signal of benzidine at 1609 cm −1 is linearly correlated with hCE1 in the concentration range of 0.1–120 ng·mL −1 . Owing to their high sensitivity and excellent SERS sensing capability, AuNPs@Cu-TCPP nanocomposites have potential applicability in the analysis of clinical samples.
研究领域:
生物医学工程
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分析化学
·
肿瘤标志物检测
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DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Guojun Weng,Xun Shen,Jianjun Li,Jingyuan Wang,Jian Zhu,Junwu Zhao
The etching of gold nanorods/nanobipyramid, or silver-coated nanorods/nanobipyramid inducing plasmon changes represents an efficient strategy to improve the performance of enzyme-linked immunosorbent assay (ELISA). However, the effect of shape on the sensitivity was negligible, especially the thickness of coated silver shell. Here, we propose a plasmonic ELISA for multi-colorimetric detection of CRP based on the etching of Ag-coated Au nanobipyramid (Au [email protected] ). The effect of silver shell thickness on the sensitivity of plasmon peak shifting was investigated by experiments and DDA calculations. The relationship between the Ag shell thickness and the sensitivity of plasmon peak shifting was obtained. Our results reveal that the thickness of coated Ag shell acts as a key factor in the multi-color change of Au [email protected] etching. It is found that Au [email protected] with medium Ag shell thickness and rod-like shape has the higher sensitivity and is suitable for sensing. At the optimized most sensitive Ag shell, the detection limit of proposed plasmonic ELISA for CRP was determined to be 0.09 ng/mL with a spectrometer in the range from 0.09 ng/mL to 25 ng/mL. Importantly, the visual detection limit was 0.78 ng/mL, which allows the differential diagnosis with the naked eye. Compared with traditional ELISA with the monochromatic intensity variations, the multi-color ELISA proposed in this study has a large linear range and rich color variation for high-sensitivity and naked-eye semi-quantitative detection.
研究领域:
纳米技术
·
生物传感
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分析化学
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材料科学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2023
·
IF 3.4
Shijie Cao,Leyao Li,Hongwei An,Guobin Mao,Junbiao Dai,Yingxin Ma
Sensitive and accurate detection of interleukin 6 (IL-6) is crucial for the early diagnosis of cerebral infarction to improve patient survival rates. However, the low-abundance of IL-6 in cerebral infarction presents a significant challenge in developing effective diagnosis method. Herein, we studied and analyzed the strong fluorescence property of 4-aminophenol phosphate (APP) and developed an enzyme-linked immunosorbent assay (ELISA) for IL-6 detection. The detection was based on the integration of optical signal change induced by alkaline phosphatase (ALP)-catalyzed APP hydrolysis and ALP-mediated ELISA. The generated colorimetric signal of 4-aminophenol, APP hydrolysis product, was used for ELISA of IL-6 with a detection limit of 0.1 ng/mL, and the visual detection of IL-6 was achieved. The changes in APP fluorescence have a good linear relationship with the logarithm of IL-6 concentration in the range of 0.005 ng/mL to 5.0 ng/mL, with a detection limit of 0.001 ng/mL, which was 100 times lower than that of conventional pNPP-based ELISA. Furthermore, the constructed ELISA effectively distinguished between samples from patients with cerebral infarction and volunteers with non-cerebral infarction, and the severity of symptoms was well distinguished based on IL-6 measurement. The dual-mode ELISA demonstrated high feasibility of low-abundance biomarker detection and displayed good potential for accurate in vitro diagnosis.
研究领域:
生物医学工程
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临床诊断
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生物化学
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2019
·Vol. 411
·期 21
·5437-5444
·
IF 3.286
Jiao Xiangyu,Zhou Yabin,Zhao Dan,Pang Duo,Wang Changtao,Du Hongwu,Wen Yongqiang,Zhang Xueji
To meet the need for high-throughput immunoassays, many multiplex fluorescent immunoassays have been proposed. Most of them need different kinds of fluorescent label indicators during the test. In this work, a novel indirect ELISA-inspired dual-channel fluorescent immunoassay based on 3-mercaptopropionic acid capped CdTe/ZnS quantum dots (QDs) was constructed. The ELISA wells were coated with two kinds of antigen–QD complex. When the primary antibodies were present in a sample, they mediated the binding of a secondary antibody–DNA–gold nanoparticle complex to the antigen–QD complex. Then the gold nanoparticles quenched the fluorescence of the QDs and a decrease in fluorescence intensity was observed. Thus, the amount of primary antibody could be estimated from the decrease of fluorescence intensity. Owing to the wide absorption range and the relatively narrow emission band of the QDs, the dual-channel fluorescent immunoassay system could work at the same excitation wavelength and the emission wavelengths of each channel had no interference. As a result, two different kinds of primary antibody could be detected at the same time in one ELISA well, which simplified the operation and greatly improved the efficiency. Besides, only one type of secondary antibody needs to be added to the prepared microtiter plates, which further simplified the operation during the detection procedure. This dual-channel fluorescent immunoassay system will provide new insights into high-throughput immunodetection.
Graphical abstract
研究领域:
生物化学
·
分析化学
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纳米技术
·
免疫学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL LETTERS
·
2024
·
IF 1.6
Haixia Yang,Xiangde Lai,Chenglong Jiang,Xuan Zhao,Huajie Pang,Dongxia Li,Zhijun Gao,Bin Qiao,Feng Han,Jia Tian
Heart failure (HF) is the leading cause of death from cardiovascular disease, and early diagnosis using serum markers holds extraordinary significance for the treatment of patients. N-terminal brain natriuretic peptide (NT-proBNP) is the gold standard for HF diagnosis. Herein, a novel, rapid, sensitive, and immune-sensing strategy for NT-proBNP is reported employing an enzyme-linked immunosorbent assay (ELISA) that combines the use of magnetic biological probes and the fluorogenic substrate Ampliflu Red. Functionalized immunomagnetic nanoparticles with large surface areas provide an abundant reaction platform and reduce the analysis time due to their superparamagnetic properties. Ampliflu Red is catalyzed by horseradish peroxidase (HRP) using hydrogen peroxide (H2O2) to produce strong fluorescence and a low background. The reported method achieved a good detection limit of 2.73 ng mL−1 and a wide linear range from 3.13 to 100 ng mL−1. The analysis time was less than 1 h, which is superior to that of traditional ELISA (3–4 h). In addition, the recoveries of spiked human serum were satisfactory, indicating that the proposed strategy has potential applications for the rapid determination of NT-proBNP and other biomarkers in clinical settings.
研究领域:
心血管疾病
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医学诊断
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生物化学
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纳米技术
·
临床化学
✓ 已验证
产品关联
DOI
高影响因子
TALANTA
·
2019
·
IF 4.916
Chen Guoning,Guo Pengqi,Wang Yan,Wang Lu,Shu Hua,Li Yunzhe,Jing Wanghui,Chang Chun,Fu Qiang
Immunoassay as a primary analytical tool is widely applied in the field of analysis. However, biological antibodies used in the routine immunoassay exhibit high cost and poor stability. Herein, in this study, a biomimetic ELISA method using molecularly imprinted polymers as a alternative to antibodies was developed. The molecularly imprinted polymers were prepared through dopamine polymerization using Fe 3 O 4 nanoparticles as magnetic nuclei, bovine serum albumin as a template, dopamine as a functional monomer and crosslinking agent. The molecularly imprinted polymers were characterized by scanning electron micrographs, transmission electron microscope, X-ray photoelectron spectroscopy, thermogravimetric analysis , vibrating sample magnetometer and X-ray diffractometer, respectively. The detection range of the established biomimetic ELISA method was 5–1000 μg mL −1 . The method exhibited high selectivity for bovine serum albumin compared with other proteins and good recovery ranging from 89.0% to 102.3% was obtained from spiked bovine serum samples. The results showed that the method by using molecularly imprinted polymers as biomimetic antibodies could be used to detect bovine serum albumin rapidly in bovine serum with a high sensitivity and accuracy. This study demonstrates that the preparation of molecularly imprinted polymers by dopamine polymerization can produce material with high affinity and potential to replace antibodies in biomimetic ELISA for protein detection.
研究领域:
分析化学
·
生物化学
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材料科学
·
生物技术
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产品关联
DOI
PubMed
高影响因子
Food Analytical Methods
·
2018
·Vol. 11
·期 9
·2553-2560
·
IF 2.245
Zhang Yuanyuan,Li Ming,Cui Yin,Hong Xia,Du Daolin
Tyramine signal amplification (TSA), the excellent signal amplification strategy, has the potential to improve the sensitivities of analytes analysis. In this work, the sensitivity of enzyme-linked immunosorbent assays (ELISA) has significantly improved by coupling TSA system and further biotin-streptavidin (BSA) system. Thus, a sensitive TSA-ELISA based on TSA was developed for detecting aflatoxin B 1 (AFB 1 ) in edible oil samples. Under optimal conditions, the limit of detection (LOD, IC 10 ) and the half-maximal inhibition concentration (IC 50 ) of the TSA-ELISA were 0.004 and 0.039 ng/mL for AFB 1 , respectively. The developed TSA-ELISA for AFB 1 has an 11-fold improved LOD value and 6-fold improved IC 50 value when compared with ELISA. The cross-reactivities of the TSA-ELISA with its analogues were negligible (< 3.48%), which indicated high specificity. The spiked recoveries were 81.4 to 118.8% with relative standard deviations (RSDs) of 3.8 to 9.0% for AFB 1 in edible oil samples. Furthermore, the results of TSA-ELISA correlated well with those obtained by HPLC-fluorescence detector. The proposed TSA-ELISA was a satisfactory tool for sensitive, inexpensive, high-throughput, and alternative detection of AFB 1 in edible oil samples. This study could provide the strategy for improving the sensitivity of ELISA with simple and practical approach, which has significant popularizing value and application prospect.
Comparison of the ELISA and TSA-ELISA for aflatoxin B 1 .
研究领域:
食品科学
·
分析化学
·
毒理学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）