JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
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2024
·
IF 3.1
Jiexia Li,Zhuqiao Fan,Hanqi Chen,Ernestina Maria Da Costa,Xiaomian Zhou,Nan Yu
Adiponectin (ADPN), which serum/plasma adiponectin levels are closely associated with insulin resistance and type 2 diabetes , and lower adiponectin levels predict an increased risk of diabetes, is a strong indicator of diabetes risk in people at high risk of diabetes in different races. Using the unique principle and performance advantages of chemiluminescence immunoassay (CLIA), an ADPN-CLIA method with high sensitivity, high specificity and wide detection range was established based on the principle of two-steps method of sandwich-type, with the magnetic particles (MPs) as the solid phase carrier and acridinium ester (AE) as the chemiluminescence reaction system. The selection of the main raw materials required, the preparation conditions of MPs-coated antibodies, the methods of AE-labeled antibodies, sample requirements and reaction modes were optimized and evaluated. AE labeling experiment was successfully performed with the labeling efficiency of 8.366 and the antibody utilization rate of 96.8%. The chemiluminescent immunoassay for ADPN had a good linear relationship from 0 ng/mL to 250 ng/mL (R 2 =0.9993), with the detection limit of 0.05 ng/mL. The coefficient of variation (CV) of intra-assay and inter-assay precision were both less than 5% respectively. The recovery rates for accuracy were from 91.26% to 107.46%. The comparison experiment of 80 clinical serum samples between the developed ADPN-CLIA with the immunoturbidimetry showed that the correlation coefficient was 0.956, and the Bland-Altman analysis showed that the limits of agreement were − 0.364 and 0.433.
研究领域:
医学检验
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生物化学
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产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2021
·Vol. 13
·期 15
·1795-1802
·
IF 2.896
Feng Wang,De-Bin Wan,Yu-Dong Shen,Yuan-Xin Tian,Zhi-Li Xiao,Zhen-Lin Xu,Jin-Yi Yang,Yuan-Ming Sun,Bruce D. Hammock,Hong Wang
The natural mycotoxin tenuazonic acid (TeA) in foods is identified as the most toxic mycotoxin among the over 70 kinds of secondary toxic metabolites produced by Alternaria alternata. Some hapten–antibody-mediated immunoassays have been developed for TeA detection in food samples, but these methods show unsatisfactory sensitivity and specificity. In this study, a rationally designed hapten for TeA mycotoxin generated with computer-assisted modeling was prepared to produce a highly specific camel polyclonal antibody, and an indirect competitive chemiluminescence enzyme immunoassay (icCLEIA) was established with a limit of detection of 0.2 ng mL−1 under optimized conditions. The cross-reactivity results showed that several analogs and some common mycotoxins had negligible recognition by the anti-TeA polyclonal antibody. The average recoveries spiked in fruit juices were determined to be 92.7% with an acceptable coefficient of variation, and good correlations between icCLEIA and liquid chromatography tandem mass spectrometry (LC-MS/MS) results were obtained in spiked samples. This developed icCLEIA for TeA detection with significantly improved sensitivity and satisfactory specificity is a promising alternative for environmental monitoring and food safety.
研究领域:
食品科学
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免疫学
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产品关联
DOI
PubMed
高影响因子
EUROPEAN FOOD RESEARCH AND TECHNOLOGY
·
2021
·Vol. 247
·期 9
·2171-2181
·
IF 2.998
Zhang Bo,Li Hongzhou,Li Yan,Fu Xiaoling,Du Daolin
Sensitive and rapid analysis of zearalenone (ZEN) in grains and feed is essential to guarantee the food security for human health. Herein a chemiluminescence method was demonstrated to measure the content of zearalenone in food samples. An immunomagnetic beads-anti-ZEN monoclonal antibody complex was prepared. Besides, the immunomagnetic beads were utilized as the solid support for rendering the immunoassay through the rapid formation of immune complexes in the solution, rapid separation and efficient heterogeneous detection. Under the optimal experimental parameters, the standard curve of ZEN detection is y = − 0.3839x + 0.1565 ( R 2 = 0.9978), the signal ratio has a well linear relationship with the logarithm of ZEN concentration in the range of 0.03–2.43 ng/mL. The limit detection of zearalenone in the developed assay was 0.01 ng/mL. The applicability of the reported approach for zearalenone detection in grains was demonstrated. In addition, the good recoveries of zearalenone were obtained, ranging from 85.15 to 102.60% at 5, 50, 150 ng/g spiked ZEN concentrations. The relative standard deviation was < 10%. The proposed chemiluminescence immunoassay is sensitive, simple, rapid and reliable for monitoring of zearalenone contamination, which provides a powerful avenue for further application of other mycotoxin in food analysis.
研究领域:
食品科学
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免疫学
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分析化学
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产品关联
DOI
高影响因子
JOURNAL OF CLINICAL LABORATORY ANALYSIS
·
2017
·Vol. 32
·期 3
·e22311
·
IF 1.521
Jingkun Wang,Yizhen Fang,Peihua Li,Huayue Lin,Hongwei Jin,Lei Gao,Shengxiang Ge,Zhong-Ying Zhang
Background To evaluate the performance of a chemiluminescence detection kit for cardiac troponin T (cTnT). Methods According to the “Guiding principles on performance analysis of diagnostic reagents in vitro” and the Clinical and Laboratory Standards Institute (CLSI) Guidelines, we assessed the detection limit, linear range, reportable range, accuracy, precision, cross reactivity, interference factors, and matrix effect of the kit, and compared these parameters with that of the commercial electrochemiluminescence detection kit for cTnT (Roche). Results The minimum detection limit of the kit was 0.01 ng/mL. The linear range was 0.01 ng/mL-25 ng/mL. The reportable range was from 0.01 ng/mL to 100 ng/mL. Precision within the batch was 2.9%-6.4%, and precision between batches was 6.0%; the accuracy was good and the recovery rate reached 96.2%. The cross-reaction test demonstrated that there was no reactivity between cTnT and a variety of troponins. Test results deviated by less than ±10% in the presence of hemoglobin ≤1000 μg/mL, bilirubin ≤250 μg/mL, triglycerides ≤11.25 mg/mL, and rheumatoid factor ≤206 U/mL, suggesting that kit results were not significantly interfered with these factors. Results from the matrix-effect assessment revealed absence of a matrix effect in the tested serum samples. Correlation study revealed that the performance of the kit was very consistent with that of the Roche electrochemiluminescence detection kit (Kappa = 0.900, P < .001) with a high correlation ( r = .903, P < .001) and a total matching rate of 95.00%. Conclusion The performance of the evaluated chemiluminescence immunoquantitation kit for cTnT detection was acceptable in all tested parameters, fulfilling requirements for clinical applications.
研究领域:
医学检验
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生物化学
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产品关联
DOI
PubMed
高影响因子
COLLOIDS AND SURFACES B-BIOINTERFACES
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2024
·
IF 5.4
Shihong Zhu,Daohang Du,Zhimin Zhao,Xianfeng Chu,Daoxiang Su,Shuli Yu,Ting Tao,Yong Jiang,Zhifei Wang
Magnetic polymer microspheres with superparamagnetism, high specificity, and monodispersity play a crucial role in the field of in vitro diagnostics. However, the surface modification process of magnetic beads is often complex, and it remains a significant challenge to prepare high-performance magnetic beads easily. To overcome these drawbacks, herein we fabricated functional interface on magnetic bead with the various amino acid via the ring-opening reaction of amino acids with epoxy groups, with attempt to produce carboxylated magnetic beads (MPS-GA) in a convenient way. Results indicate that when compared to other amino acids, the phenylalanine magnetic beads (MPS-GA1) developed in this study exhibit strong adsorption for mouse immunoglobulin (IgG), streptavidin (SA), and protamine (PA), with an IgG adsorption capacity of 53.5 μg/mg and a coupling capacity of 52.5 μg/mg. It is found that electrostatic forces and hydrophobic interactions are key factors influencing biomolecular interactions. Additionally, these magnetic beads can generate strong chemiluminescent signals, significantly reducing background levels by up to 99.7 %. Therefore, the magnetic beads proposed in this paper can serve as carriers for chemiluminescent immunoassay (CLIA), providing new insights into the synthesis of high-quality magnetic bead.
研究领域:
生物医学工程
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诊断学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Dezhi Huang,Enqi Huang,Dongyang Cai,Zhenhua Chen,Hongting Wen,Yu Wang,Dachuan Ma,Yao Lu,Xianming Liu,Dayu Liu
Digital microfluidics (DMF) features programmed manipulation of fluids in multiple steps, making it a valuable tool for sample pretreatment. However, the integration of sample pretreatment with its downstream reaction and detection requires transferring droplets from the DMF device to the outside world. To address this issue, the present study developed a modified DMF device that allows automated droplet ejection out of the chip, facilitated by a tailor-designed interface. A double-layered DMF microchip with an oil-filled medium was flipped over, with a liquid infusion port and a liquid expulsion port accommodated on the top working PCB plate and the bottom grounded ITO plate, respectively, to facilitate chip-to-world delivery of droplets. Using chemiluminescent immunoassay (CLIA) as an illustrative application, the sample pretreatment was programmed on the DMF device, and CLIA droplets were ejected from the chip for signal reading. In our workflow, CLIA droplets can be ejected from the DMF device through the chip-to-world interface, freeing up otherwise occupied electrodes for more sample pretreatment and enabling streamlined droplet microreactions and batch-mode operation for bioanalysis. Integrated with these interfacing portals, the DMF system achieved a single-channel throughput of 17 samples per hour, which can be further upscaled for more productive applications by parallelizing the DMF modules. The results of this study demonstrate that the droplet ejection function that is innovated in a DMF sample pretreatment microsystem can significantly improve analytical throughput, providing an approach to establishing an automated but decentralized biochemical sample preparation workstation for large-scale and continuous bioanalysis.
研究领域:
生物分析
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医学检测
·
微流控技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2018
·
IF 6.042
Yuchen Jin,Qi Kang,Xinli Guo,Bin Zhang,Dazhong Shen,Guizheng Zou
Signal amplification for electrochemiluminescence (ECL) has conventionally been achieved by employing effective matrixes that can accelerate the electrochemical redox processes or carry more electrochemiluminophores. Herein, a convenient signal-amplification strategy was proposed for an ECL immunoassay with carboxylated g-C3N4 nanosheets (NSs) as tags and carcinoembryonic antigen (CEA) as the model target via electrochemically pretreating the substrate: a glassy-carbon electrode (GCE) modified with a polymerized 2-aminoterephthalic acid (ATA) film (GCE/ATA). Bioconjugates of g-C3N4 NSs and the signal CEA antibody (Ab2) (i.e., g-C3N4 NS–Ab2) were immobilized on GCE/ATA via a sandwich immunoreaction to form GCE/ATA–Ab1–Ag–Ab2–NSs. Electrochemical-impedance spectroscopy and potential-resolved ECL characterization proved that GCE/ATA plays an important role in the electron-transfer resistance (Ret) of the GCE/ATA–Ab1–Ag–Ab2–NSs for ECL and that successively scanning GCE/ATA–Ab1–Ag–Ab2–NSs from 0 to −1.6 V in K2S2O8- and H2O2-containing medium could reduce the Ret and bring out 3.3-times-enhanced ECL at the 10th scan cycle compared with that of the 1st scan cycle, which was about 10.2 times the ECL of the GCE/ATA–Ab1–Ag–Ab2–NSs in medium containing merely K2S2O8. Inspired by this, direct and successive scanning of GCE/ATA in K2S2O8- and H2O2-containing medium was employed during fabrication, which dramatically reduced the Ret of GCE/ATA–Ab1–Ag–Ab2–NSs and brought out obviously enhanced ECL responses for selectively determining CEA from 0.1 pg/mL to 1 ng/mL, with a detection limit of 3 fg/mL.
研究领域:
分析化学
·
生物医学工程
✓ 已验证
DOI
PubMed
高影响因子
TALANTA
·
2024
·
IF 5.6
Danyi Zhao,Mai Hu,Chenghao Hu,Di Wang,Hailun Chen,Yangwei Ou,Rongli Liu,Xiaoyang Li,Long Wu,Peng Liu,Zhiwei Shen,Qi Chen
Microcystin-LR (MC-LR), a toxic cyanobacterial toxin in freshwater, poses significant health and ecological risks due to its ability to induce cell apoptosis and liver damage. Sensitive detection of MC-LR is crucial for public health and water safety. In this work, we engineered a multivalent bifunctional nanobody (A2.3-C4-SBP) by fusing the anti-MC-LR nanobody gene (A2.3) with self-assembling peptides (C4) and a streptavidin-binding peptide (SBP). A2.3-C4-SBP was directionally immobilized on the ELISA microplate via streptavidin-mediated to develop a multivalent bifunctional nanobody-based chemiluminescent immunoassay (MBN-CLIA) for MC-LR detection in lake water. The IC 50 of the A2.3-C4-SBP heptamer based CLIA was 5.80 ng/mL, and the LOD (IC 10 ) was 0.33 ng/mL, which were 9.51-fold and 1.82-fold lower, respectively, than those of the A2.3-SBP monomer based CLIA. Additionally, the IC 50 and LOD were 1.26-fold and 1.82-fold lower, respectively, than those of the A2.3-C4-SBP heptamer without streptavidin-mediated directional immobilization. In summary, this work developed a sensitive, rapid and simple immunoassay for the detection of MC-LR in lake water based on multivalent bifunctional nanobodies. Furthermore, the proposed combined strategy of nanobody multimerization and directed immobilization is simple to operate and has great potential to improve the sensitivity and signal amplification of various immunoassays.
研究领域:
环境科学
·
免疫学
·
生物技术
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 9
·1-8
·
IF 5.479
Fang Dandan,Zhang Shupei,Dai Hong,Li Xiuhua,Hong Zhensheng,Lin Yanyu
An ultrasensitive competitive-type electrochemiluminescence immunoassay for the mycotoxin zearalenone is described. The method is based on the use of (a) a mimotope peptide that was selected from a phage displayed peptide library and used to substitute ZEN for designing the competitive assay; (b) NiFe 2 O 4 nanotubes with large specific surface area loaded with the ECL probe Ru(bpy) 3 2+ ; and (c) poly(vinylpyrrolidone) (PVP)-assisted synthesis of TiO 2 mesocrystals that acts as the sensing platform and support for antibody immobilization. Under the optimized conditions and at an ECL working potential of 1.1 V, a linear response is found for ZEN in the 0.1 to 1.0 × 10 −5 ng·mL −1 concentration range with a detection limit as low as 3.3 fg·mL −1 .
An ultrasensitive competitive-type electrochemiluminescence (ECL) immunosensor based on mimotope peptide was constructed for the detection of Zearalenone.
研究领域:
食品科学
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分析化学
·
毒理学
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2021
·
IF 6.558
Huan Zhao,Xinghan Qiu,Enben Su,Li Huang,Yunfeng Zai,Yuan Liu,Hui Chen,Zunliang Wang,Zhu Chen,Song Li,Lian Jin,Yan Deng,Nongyue He
The concentration ratio of glycosylated hemoglobin A1c (HbA1c) to total hemoglobin (Hb) has long been used to accurately determine stagewise diabetes because this parameter represents a reliable and accurate biomarker of mean 90-day blood glucose values. In this paper, we report a time-resolved chemiluminescence assay that can detect both Hb and HbA1c. For the determination of Hb, the interaction of heme in Hb with H 2 O 2 in NaOH solution was performed to generate a chemiluminescence peak. HbA1c was detected using a sandwich immunoassay based on an acridine ester-labeling method using the same Hb chemiluminescence trigger system. The results showed that the repeatability %CV of the proposed method for multiple detections of HbA1c and Hb ranged from 1.22 to 2.21%, with a median value of 1.73%, while the within-site reproducibility %CV ranged from 2.13 to 3.27%, with a median value of 2.81%. Compared with the conventional HPLC method (BIO-RAD D10 system), the correlation coefficient was 0.9959. In conclusion, a time-resolved multiple chemiluminescence immunoassay biosensor for HbA1c/Hb detection was established, and the method has excellent reproducibility and accuracy, thus demonstrating great potential for clinical application.
研究领域:
医学诊断
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生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF ELECTROANALYTICAL CHEMISTRY
·
2016
·
IF 2.822
Zheng-Jun Huang,Wen-Di Han,Yan-Hong Wu,Xiang-Guang Hu,Ya-Ni Yuan,Wei Chen,Hua-Ping Peng,Ai-Lin Liu,Xin-Hua Lin
A versatile electrochemiluminescent (ECL) immunoassay based on the use of magnetic nanoparticles (MNPs) that act as the carrier for the immobilization of the primary antibody (Ab 1 ) against α-fetoprotein (AFP) has been developed. CdTe quantum dots (QDs) were used to label the same antibody to give a secondary antibody (Ab 2) for use in a sandwich immunoassay. On formation of the MNP-Ab 1 /AFP/QD-Ab 2 sandwich immunocomplex, a strong ECL signal has been obtained at a peak potential of − 1.69 V. The principle of the ECL detection for target AFP was on account of the increment concentration of QD-Ab 2 conjugates after immunoreaction, which resulted in the increase in ECL intensity. The linear response range was between 1.0 and 200 ng mL − 1 , and the detection limit was 0.32 ng mL − 1 for the detection model of AFP. The assay was applied to the determination of AFP in serum by the standard addition method. The immunoassay presented here allows for easy separation and purification of AFP, exhibit strong ECL, highly sensitive, well reproducible and precise. In our preception, it has great potential for detecting other proteinic tumor biomarkers.
研究领域:
生物医学工程
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS
·
2024
·
IF 4.9
Daoxiang Su,Jiaxing Han,Lucheng Chen,Shihong Zhu,Yong Jiang,Zhifei Wang
Inhibiting the nonspecific protein adsorption on the surface of magnetic microspheres has become a pressing challenge for immunoassays . In this paper, a novel dextran-modified sandwich-type magnetic polymer microsphere was developed and demonstrated to be a promising carrier for chemiluminescence immunoassay (CLIA). Dextran-modified sandwich-type magnetic polymer microspheres (PFS-PGDN) were synthesized by surface-initiated atom transfer radical polymerization (SI-ATRP) and ring-opening reaction of epoxy and aminodextran on the surface of magnetic P(GMA-MMA)-NH 2 @Fe 3 O 4 @SiO 2 (PFS) microspheres . Then, carboxylated dextran-modified sandwich-type magnetic polymer microspheres (PFS-PGDC) were prepared by the surface carboxylation of the microspheres, which allowed the prepared microspheres to better couple with nucleic acids or antibodies. It is found that after grafting dextran , magnetic PFS-PGDC microspheres could significantly reduce the nonspecific protein adsorption by over 90 %. The chemiluminescence immunoassay assay showed a reduction of the background value to a very low level, indicating that the magnetic spheres showed significant advantages in improving the sensitivity of the chemiluminescence assay. Moreover, the chemiluminescence sensitivity and signal values of the synthesized magnetic microspheres have reached high levels. Therefore, the magnetic microspheres synthesized here hold great promise for applications in the field of CLIA.
研究领域:
生物材料
·
免疫学
·
分析化学
✓ 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）