Food Bioscience
·
2024
·
IF 4.8
Jiao Li,Panpan Cui,Zhijia Wang,Yang Lu,Yuan Ding,Xiude Hua
Haloxyfop-P-methyl, a widely used aryloxyphenoxypropionate herbicide, has been identified as a potential hazard to ecological security and human health. Consequently, it is essential to develop superior methods for detecting haloxyfop-P-methyl. In this study, 12 peptidomimetics and 4 anti-immunocomplex peptides were isolated from phage-displayed peptide libraries, which can specifically recognize the anti-haloxyfop-P-methyl antibody and the immunocomplex formed by haloxyfop-P-methyl and its antibody, respectively. Following the sensitivity assessment of each phage-displayed peptide, competitive and noncompetitive phage enzyme-linked immunosorbent assays (P-ELISAs) were developed using the best phage-displayed peptides. The half inhibition concentration (IC 50 ) and half saturation concentration (SC 50 ) of competitive and noncompetitive P-ELISAs were determined to be 8.15 and 1.83 ng/mL, respectively, representing the 5-fold and 22-fold increase in sensitivity compared to previously reported immunoassay. The competitive P-ELISA exhibited 2.2%–114.8% cross-reactivities (CRs) to haloxyfop-methyl, haloxyfop and quizalofop-p-ethyl, while the noncompetitive P-ELISA showed no significant CR (<0.1%) except for haloxyfop-methyl (CR = 41.6%). The average recoveries of two P-ELISAs in soil, soybean, potato, and cabbage samples ranged from 78.4% to 104.3% and 76.8%–95.2%, respectively. Additionally, the results of P-ELISAs for blind samples exhibited a good correlation with those obtained from ultra-performance liquid chromatography/tandem mass spectrometry.
研究领域:
环境科学
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分析化学
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毒理学
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DOI
高影响因子
Drug Testing and Analysis
·
2014
·Vol. 6
·期 10
·1020-1027
·
IF 2.84
Yajing Lei,Qian Zhang,Lizheng Fang,Muhammad Sajid Hamid Akash,Kanwal Rehman,Zhiming Liu,Weixing Shi,Shuqing Chen
We simultaneously set up two competitive (direct and indirect) enzyme-linked immunosorbent assays (ELISAs) based on the same antibody for estimation of cotinine (COT) in pregnant women especially and population generally exposed to environmental tobacco smoke. The results show that the limits of detection (LODs) for direct competitive ELISA and indirect competitive ELISA were 0.04 μgL -1 and 0.1 μgL -1 , respectively. Direct competitive ELISA was found to be more sensitive than indirect competitive ELISA. Thereafter, we applied our direct competitive ELISA for the detection of COT from urinary samples taken from 450 volunteers from the Zhejiang Province of China. COT was detected in 100% of participants with concentration ranging from LOD to 5358.0 μgL -1 . The GM and 95th percentile concentration of COT in pregnant women were 6.3 μgL -1 and 57.2 μgL -1 , respectively. Males had statistically higher COT concentrations than females ( P < 0.0001), active smokers had statistically higher COT concentrations than non-smokers ( P < 0.0001), whereas, non-pregnant women were found to have higher COT concentration than pregnant women. We conclude that our developed direct competitive ELISA is useful for detecting the COT in urinary concentration of human. The human urinary data obtained in this study indicated that common people generally and pregnant women especially were highly exposed to COT. Further studies are needed to focus on the sources of exposure, potential health effects and risk assessment of exposure to COT. Copyright © 2014 John Wiley & Sons, Ltd.
研究领域:
分析化学
·
环境卫生
·
公共卫生
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2017
·
IF 7.78
Yang Liu,Meng Cai,Weiping Wu,Yuting Fang,Pei She,Shouming Xu,Jianguo Li,Kang Zhao,Jingjuan Xu,Ning Bao,Anping Deng
Herein, we reported a disposable electroanalytical device for competitive enzyme-linked immunosorbent assay (ELISA) of phenylethanolamine A (PA). Conductive carbon tape coated with gold cluster served as the working electrode , while the counter and reference electrodes were fabricated on the filter paper by the screen-printing technique. Separating fabrication of the working electrode from other electrodes could make it possible for full utilization of the working electrode modification in bulk and no contamination of the immune-reagents on the counter and reference electrodes. The gold cluster played an important role in both immobilizing antigen and accelerating electron transfer . The eight-channel devices were utilized to detect PA on the strategy of competitive ELISA. The detection range and the limit of detection (LOD) using differential pulse voltammetry for PA were 0.005–60 ng mL −1 and 2.6 pg mL −1 , respectively. And also, the linear range for PA performed by square wave voltammetry was 0.05–60 ng mL −1 with the LOD value as 0.028 ng mL −1 . The results clearly demonstrated that the proposed electroanalytical devices could be successfully applied in immunoassay and might be further developed for determination of different analytes based on ELISA format.
研究领域:
分析化学
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免疫学
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DOI
PubMed
高影响因子
Analytical Methods
·
2021
·Vol. 13
·期 46
·5542-5548
·
IF 2.896
Xinfa Cai,Meijuan Liang,Fei Ma,Sherif Ramzy Mohamed,Amira Abdel Goda,Dawood H. Dawood,Li Yu,Peiwu Li
A direct competitive nanozyme-linked immunosorbent assay (dcNLISA) based on MnO2 nanosheets (MnO2 NSs) as a nanozyme label was developed for the highly sensitive determination of fumonisin B1 (FB1). MnO2 NS-labeled fumonisin B1-bovine serum albumin was easily synthesized as a competing antigen for the dcNLISA. And color changes derived from the MnO2–3,3′,5,5′-tetramethylbenzidine (TMB) system were exploited as the output signals of the dcNLISA. Several experimental parameters including the concentrations of the coating antibody, pH values, ionic strength and methanol concentration were optimized. Under the optimal conditions, the proposed method demonstrated a linear range (1.17–20.74 ng mL−1) with a reliable correlation coefficient (R2 = 0.9989), a satisfactory limit of detection (0.63 ng mL−1) and high selectivity for the detection of FB1. The recoveries of FB1 in spiked corn and wheat samples were in the range of 85.31–108.16% with coefficients of variation (CVs) ranging from 6.14% to 9.23%. Meanwhile, the testing results showed good consistency (R2 = 0.9892) between the developed dcNLISA and the reference method, liquid chromatography/mass spectrometry/mass spectrometry (LC-MS/MS) method. The proposed method was proven to be simple, sensitive, cost-effective and reliable for the screening of FB1 in cereals.
研究领域:
食品科学
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分析化学
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生物技术
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2023
·
IF 6.1
Chang Liu,Hong Lin,Limin Cao,Kaiqiang Wang,Jianxin Sui
The third generation of genetic engineering antibodies, single-domain antibodies, have been widely reported as potential biomaterials in recognizing small molecular hazards. In this study, a shark-derived single-domain antibody was used as the recognition element for the first time to detect enrofloxacin (ENR), one of the most representative hazards in aquaculture. An ENR-specific clone named 2E6 was isolated by phage display technology. Experimental results proved that 2E6 ssdAb showed high affinity to ENR-PEI complete antigen, with the highest OD 450 value of 1.348 in binding ELISA . Through icELISA, it was determined that the IC 50 of 2E6 ssdAb to ENR was 19.230 ng/mL, while the IC 10 was 0.975 ng/mL, with rare recognition to other fluoroquinolones, which showed high sensitivity and specificity to ENR. The 2E6 ssdAb also performed excellently in fish matrix immunoassay . Results showed that the ENR-negative fish matrix did not seriously interfere with the recognition of 2E6 ssdAb to ENR-OVA, with the matrix index between 4.85% and 11.75%, while the results of icELISA in ENR-spiked fish matrix showed that 2E6 ssdAb could recognize the target ENR in different ENR-spiked concentrations of the fish matrix (10–1000 ng/mL), with the recovery between 89.30% and 126.38% and the RSD between 1.95% and 9.83%. This study broadens the application scenario of shark-derived single-domain antibodies as small molecule recognition biomaterials, providing a new recognition element on ENR detection for immunoassay.
研究领域:
分子生物学
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免疫学
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水产养殖
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生物技术
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DOI
PubMed
高影响因子
JOURNAL OF FOOD COMPOSITION AND ANALYSIS
·
2025
·
IF 4.6
Wenjing Liu,Guangjie Wu,Liping Wang,Jiaying Zhao,Renrong Liu
Immunoaffinity column (IAC) cleanup, coupled with high performance liquid chromatography (HPLC) analysis, is one of the most reliable methods for aflatoxin B1 (AFB1) analysis. However, real-time monitoring of the IAC capacity remains unachievable in IAC-HPLC analysis, compromising the reliability of AFB₁ quantification. Two artificial structural analogues of aflatoxin (AFB1-OH and AFB2-OH) were synthesized as potential markers for IAC capacity monitoring. Competitive ELISA and recovery experiments demonstrated that AFB2-OH was more suitable as a marker. When AFB2-OH was used for real-time monitoring of the capacity of the IAC in AFB1 determination by HPLC, the recovery of AFB1 exceeded 95 %. The limit of detection (LOD) and the limit of quantification (LOQ) for AFB1 were determined to be 0.14 ng/mL and 0.46 ng/mL, respectively. The method also exhibited good precision and accuracy. In spiked recovery experiments of four grains, the recovery of AFB1 ranged from 85.16 % to 107.05 % with relative standard deviations of 0.89 %-4.98 %. More importantly, due to the addition of AFB2-OH, the capacity of the IAC was monitored in real-time, ensuring the reliability of AFB1 determination results.
研究领域:
分析化学
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食品安全
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生物化学
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色谱分析
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免疫分析
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DOI
高影响因子
MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 11
·1-12
·
IF 6.232
Yang Huiyi,He Qiyi,Chen Yingshan,Shen Ding,Xiao Huanxin,Eremin Sergei A.,Cui Xiping,Zhao Suqing
Platinum nanoflowers (PtNFs) were utilized in a competitive enzyme-linked immunosorbent assay (ELISA) and in a lateral flow immunoassay (LFIA) for superior peroxidase-like activity and intense brown color, respectively. PtNFs were linked to the polyclonal antibody (pAb) to form the pAb-PtNFs probes for the dual immunoassay. Based on optimized pAb-PtNF probes, both enzyme-linked immunosorbent assay (PtNFs-ELISA) and lateral flow immunoassay (PtNFs-LFIA) perform very well. The absorbance at 450 nm decreases linearly in the DHEA concentration range 2.1 to 118.1 ng mL −1 , and the limit of detection is 1.3 ng mL −1 and the IC 50 value is 15.7 ng mL −1 of PtNFs-ELISA. The visual cut-off value of PtNFs-LFIA is 10.0 ng mL −1 . The average recoveries from spiked samples range from 95.0 to 108.9% with a coefficient of variation below 12.2%. Excellent recoveries and correlation between the two methods were observed. Furthermore, the designed immunosensors exhibited good selectivity, confirming a broad development prospect in DHEA monitoring.
Graphical Abstract
研究领域:
纳米技术
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生物化学
·
分析化学
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免疫学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2023
·Vol. 415
·期 10
·1877-1887
·
IF 4.3
Wang Yinghui,Yu Ling,Zhang Hui,Zhu Runzhi,Meng Zhe
An accurate and sensitive competitive enzyme-linked immunosorbent assay (ELISA) based on persistent luminescence nanoparticles Zn 2 GeO 4 :Mn 2+ , Eu 3+ (ZGME) was developed for detecting ochratoxin A (OTA), a powerfully toxic mycotoxin usually found in grains. As a signal output element of autofluorescence-free biosensors, ZGME can be integrated into ELISA with glucose oxidase (GOx)-binding OTA molecules due to its excellent pH-responsive persistent luminescence. In the absence of OTA, the OTA-GOx conjugate was captured by the anti-OTA monoclonal antibody (anti-OTA mAb) pre-coated on the 96-well plate. The results indicate a decrease in the pH value of the solution, which triggered the quenching of ZGME luminescence due to GOx-dependent gluconic acid production. The presence of OTA inhibited the binding of OTA-GOx on the plate, thus decreasing the production of gluconic acid and increasing the persistent luminous intensity of ZGME. Under the optimized concentrations of anti-OTA mAb and OTA-GOx, quantitative determination of OTA was achieved by plotting the increase or decrease in persistent luminescence intensity of ZGME at 535 nm. In this study, the linear range was from 0.1 μg L −1 to 63 μg L −1 , and the limit of detection (LOD) was as low as 0.045 μg L −1 . In five food samples (corn grit, brown rice, soybean, rice, and wheat), the results exhibited good stability and repeatability, with a recovery range from 81.3% to 94.4% and a relative standard deviation (RSD) of less than 4.2%. Hence, the established method provides a sensitive, accurate, and autofluorescence-free approach for the determination of OTA in different grain samples.
研究领域:
分析化学
·
毒理学
·
食品科学
·
纳米技术
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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）