ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2024
·Vol. 416
·期 12
·2995-3006
·
IF 3.8
Guan Tian,Chen Sha,Chen Yanhong,Wang Yu,Yu Xiaoqin,Huang Xinan,Liu Qing,Lei Hongtao,Yang Qunhua
Stimulant laxatives were recently found to be abused in slimming foods, resulting in harmful effects on consumers. To ensure the safety of relative products, sensitive yet multiplex immunoassays are crucial in rapid screening of stimulant laxatives. However, there are few immunoassays for these substances, and even less for broad-specific recognition. Thus, in this work, four theoretically promising haptens of emerging stimulant laxative bisacodyl were rationally designed using molecular modeling and synthesized to immune animals, whose feasibility was confirmed by the obtained broad-specific antibody. Based on this unique antibody, a highly sensitive multiplex competitive indirect enzyme-linked immunosorbent assay (ciELISA) was established with low limits of detection for bisacodyl, sodium picosulfate, and BHPM (0.23, 13.68, and 0.11 ng/mL). In spiked sample recovery test and real sample detection, this ciELISA exhibited acceptable consistency with the validation method, demonstrating high accuracy and applicability of our method. This reliable multiplex ciELISA proceeds the rapid screening of stimulant laxatives in slimming foods. Graphical abstract
研究领域:
食品安全
·
免疫学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CONTROL
·
2017
·
IF 3.496
Xiya Zhang,Meirong Song,Xuezhi Yu,Zhanhui Wang,Yuebin Ke,Haiyang Jiang,Jiancheng Li,Jianzhong Shen,Kai Wen
To reduce the incidence of false-positive and false-negative results caused by high or low cross-reactivity (CR%) values of the antibodies for total aflatoxins (AFs, AFB 1 +AFB 2 +AFG 1 +AFG 2 ) detection, a new broad-specific monoclonal antibody (MAb) with uniform affinity, named 5H3, was developed. Moreover, magnetic beads (MBs) replaced microplates as immobile phase to improve the sensitivity of the enzymatic immunoassay. Then, a direct competitive enzyme-linked immunosorbent assay (ELISA) based on MBs (MBs-dcELISA) that could simultaneously detect the total AFs with similar sensitivity was developed. Following optimization of conditions, the half maximal inhibitory concentrations (IC 50 ) of the MBs-dcELISA in buffer were 0.05 ng/mL for AFB 1 , 0.04 ng/mL for AFB 2 , 0.05 ng/mL for AFG 1 , 0.06 ng/mL for AFG 2 . The corresponding CR% values were 100%, 125%, 100% and 83.3%, respectively. The limit of detection (LOD) of the MBs-dcELISA for the total AFs was 0.21 ng/g with a working range from 0.22 ng/g to 19.8 ng/g, and the recoveries for the total AFs ranged from 74.5% to 96.5% with coefficients of variation (CV) under 12.1% in spiked maize samples. In addition, the MBs-dcELISA was more sensitive than the conventional dcELISA. Finally, the MBs-dcELISA was applied to screen 9 naturally contaminated maize samples and 6 spiked samples and the results indicated a good agreement with that obtain by HPLC-MS/MS method.
研究领域:
食品科学
·
免疫学
✓ 已验证
DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2013
·
IF 5.695
Xiaodong Bi,Zhen Liu
Molecularly imprinted polymers (MIPs), as inexpensive and stable substitutes of antibodies, have shown great promise in immunoassays. Glycoproteins are of significant diagnostic value. To facilitate the application of MIPs in clinical diagnostics, a general and facile imprinting method toward glycoproteins oriented for an enzyme-linked immunosorbent assay (ELISA) in the form of a 96-well microplate is essential but has not been fully explored yet. In this study, a new method called boronate affinity-based oriented surface imprinting was proposed for facile preparation of glycoprotein-imprinted microplates. A template glycoprotein was first immobilized by a boronic acid-modified microplate through boronate affinity binding, and then, a thin layer of polyaniline was formed to cover the microplate surface via in-water self-copolymerization. After the template was removed by an acidic solution, 3D cavities that can rebind the template were fabricated on the microplate surface. Using horseradish peroxidase (HRP) as a model target, the effects of imprinting conditions as well as the properties and performance of the prepared MIPs were investigated. α-Fetoprotein (AFP)-imprinted microplate was then prepared, and thereby, a MIP-based ELISA method was established. The prepared MIPs exhibited several highly favorable features, including excellent specificity, widely applicable binding pH, superb tolerance for interference, high binding strength, fast equilibrium kinetics, and reusability. The MIP-based ELISA method was finally applied to the analysis of AFP in human serum. The result was in good agreement with that by radioimmunoassay, showing a promising prospect of the proposed method in clinical diagnostics.
研究领域:
生物化学
·
医学诊断
·
材料科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
·
2015
·
IF 0.986
Huijuan Yan,Liqiang Liu,Naifeng Xu,Hua Kuang,Chuanlai Xu
In the present study, a class-selective monoclonal antibody (mAb) for carbamate fungicides was obtained using two haptens. One hapten was designed as immunogen and the other one for coating. Under the optimum assay conditions of coating antigens H2-ovalbumin, in the assay buffer of sodium chloride 1.6%, no methanol, and pH 6.5, the assay system was more sensitive. The linear range was from 0.06 to 7.47 ng/mL. The cross-reactivity test was carried out with IC50 values of 0.45 ng/mL for carbendazim, 0.57 ng/mL for benomyl, and 9.70 ng/mL for thiabendazole. Other related compounds thiophanate methyl, fenbendazole, mebendazole, methyl paraoxon, methyl parathion, chlorpyrifos, and chlorothalonil did not cross-react significantly in this assay. To evaluate the proposed immunoassay, real samples such as apples, tomatoes, and cucumbers were subjected to recovery test with carbendazim. The recovery and coefficient variation range from 73% to 125% and 0.36% to 11.80%, respectively. The developed method was also compared with the commercial kit for assaying carbendazim in the fruits and vegetables and the results were consistent. All results indicated that this developed enzyme-linked immunosorbent assay (ELISA) is useful for the detection of carbendazim residues in fruits and vegetables.
研究领域:
生物化学
·
食品安全
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
FISH & SHELLFISH IMMUNOLOGY
·
2018
·
IF 3.185
Liting Wu,Shengli Fu,Xiaoxue Yin,Wenna Leng,Zheng Guo,Anli Wang,Jianmin Ye
Affinity maturation of the antibody response , a process of antibody affinity increasing over response, is one of the key features of the mammalian immune system. However, the process is incompletely understood in teleost, including channel catfish ( Ictalurus punctaus ). In this study, IgM affinity maturation in channel catfish was investigated by estimating the kinetics of antibody affinity using ELISA and ELISPOT assays . Fish were immunized with a T-cell dependent antigen (TNP-KLH), and individual serum IgM antibody titers and affinities, and IgM + antibody-secreting cells (ASCs) in peripheral blood were analyzed over a period of 14 weeks. A detectable serum anti-TNP response developed by 2-weeks post-immunization, and the maximal antibody production was observed by 6-weeks post-immunization. The average affinity of anti-TNP serum antibody increased consistently and reached the maximum by 10-weeks post-immunization. The increase of antibody affinity beyond the point of optimal antibody titer revealed that the affinity maturation of IgM antibody response occurred in channel catfish. Dissection of dynamics of individual affinity subpopulations indicated that a significant proportion of low affinity subpopulations appeared at early response, and high affinity subpopulations appeared predominantly at later, resulting in a 100-fold increase in affinity over response. Additional, TNP + IgM + ASCs was detected by 2-weeks post-immunization and achieved the maximal number by 6-weeks post-immunization. Using an inhibition ELISPOT assay, the findings of a consistent increase in the average affinity of secreted IgM antibody by peripheral blood ASCs, as the immune response progressed, confirmed the occurrence of the affinity maturation. Taken together, the results of this study indicated that affinity maturation occurred in channel catfish following immunization with a TD antigen TNP-KLH.
研究领域:
免疫学
·
鱼类生物学
✓ 已验证
DOI
PubMed
高影响因子
International Journal of Environmental Research and Public Health
·
2013
·Vol. 10
·期 4
·1598-1608
·
IF 1.998
Hua Kuang,Wenbing Wang,Liguang Xu,Wei Ma,Liqiang Liu,Libing Wang,Chuanlai Xu
A sensitive and specific monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (ELISA) was established and validated for the detection of staphylococcal enterotoxin A (SEA). After routine fusion and selection, 10 monoclonal antibodies showed high affinity for SEA. An optimal pair for sandwich ELISA was selected by pairwise interaction analysis. After optimization, the limit of detection (LOD) and linear dynamic range of the method were established, and were found to be 0.0282 ng/mL and 0.06–2 ng/mL, respectively. The recovery in pure milk ranged from 82.67% to 111.95% and the intra- and inter-assay coefficients of variation ranged from 3.16% to 6.05% and from 5.16% to 10.79%, respectively. Cross-reactivity with staphylococcal enterotoxin B (SEB), staphylococcal enterotoxin C (SEC), staphylococcal enterotoxin D (SED), and staphylococcal enterotoxin E (SEE) in this method were insignificant. These results indicate that the sandwich ELISA method developed in our study is effective for routine identification of SEA in food samples.
研究领域:
食品科学
·
免疫学
✓ 已验证
DOI
高影响因子
Journal of Pharmaceutical Analysis
·
2021
·
IF 4.769
Min Pei,Tingting Liu,Lu Ouyang,Jianhua Sun,Xiaojie Deng,Xiaomin Sun,Wei Wu,Peng Huang,Yi-Li Chen,Xiaorong Tan,Xiaoyue Liu,Peng Zhu,Yongzhen Liu,Deheng Wang,Junliang Wu,Qi Wang,Guifeng Wang,Likun Gong,Qiuping Qin,Chunhe Wang
Antibody-drug conjugates (ADCs) are commonly heterogeneous and require extensive assessment of exposure-efficacy and exposure-safety relationships in preclinical and clinical studies. In this study, we report the generation of a monoclonal antibody against monomethyl auristatin E (MMAE) and the development, validation, and application of sensitive and high-throughput enzyme-linked immunosorbent assays (ELISA) to measure the concentrations of MMAE-conjugated ADCs and total antibodies (tAb, antibodies in ADC plus unconjugated antibodies) in cynomolgus monkey sera. These assays were successfully applied to in vitro plasma stability and pharmacokinetic (PK) studies of SMADC001, an MMAE-conjugated ADC against trophoblast cell surface antigen 2 (TROP-2). The plasma stability of SMADC001 was better than that of similar ADCs coupled with PEG4-Val-Cit, Lys (m-dPEG24)-Cit, and Val-Cit linkers. The developed ELISA methods for the calibration standards of ADC and tAb revealed a correlation between serum concentrations and the OD 450 values, with R 2 at 1.000, and the dynamic range was 0.3–35.0 ng/mL and 0.2–22.0 ng/mL, respectively; the intra- and inter-assay accuracy bias% ranged from −12.2% to −5.2%, precision ranged from −12.4% to −1.4%, and the relative standard deviation (RSD) was less than 6.6% and 8.7%, respectively. The total error was less than 20.4%. The development and validation steps of these two assays met the acceptance criteria for all addressed validation parameters, which suggested that these can be applied to quantify MMAE-conjugated ADCs, as well as in PK studies. Furthermore, these assays can be easily adopted for development of other similar immunoassays.
研究领域:
药理学
·
免疫学
·
生物化学
·
药物开发
✓ 已验证
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2016
·
IF 7.476
Huifang Zhang,Xiaoming Ma,Shuisheng Hu,Yue Lin,Longhua Guo,Bin Qiu,Zhenyu Lin,Guonan Chen
Development of convenient but sensitive method for influenza detection is highly important in immediate and effective clinical treatment. In this study, an ultrasensitive colorimetric approach combining the advantages of the convenience of the enzyme-induced metallization and the high specificity of enzyme-linked immunosorbent assay for the detection of influenza virus A (H7N9 as model) has been developed. Two rounds of amplification are utilized to enhance the detection sensitivity. The amplification of enzymatic reaction combines with the specific optical properties of gold nanoparticles causing the enhancing of the optical signal immensely. In addition, the increased surface area and the magnetic enrichment effect also enable the magnetic bead (MB) to catch a large number of alkaline phosphatase (ALP) and detection antibody (Ab 2 ), thus very small amounts of the virus can be easily detected. Compared with conventional method, this approach exhibits outstanding sensitivity for ALP detection, 0.2 U/L of ALP can be distinguished with a spectrometer and 2 U/L with the naked eye. And as low as 25 pg/mL of H7N9 can be detected by the naked eye. This approach shows an extensive horizon for bioassays and is available in clinical diagnosis with the advances of simplification, effectiveness, low cost and sensitive readout.
研究领域:
病毒学
·
诊断医学
·
纳米技术
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2024
·
IF 10.7
Wenjing Xue,Li Wang,Kexin Yi,Lingyu Sun,Haozhen Ren,Feika Bian
Hepatocellular carcinoma (HCC), as one of the most lethal cancers, significantly impacts human health. Attempts in this area tends to develop novel technologies with sensitive and multiplexed detection properties for early diagnosis. Here, we present novel hydrogel photonic crystal (PhC) barcodes with tyramine deposition amplified enzyme-linked immunosorbent assay (ELISA) for highly sensitive and multiplexed HCC biomarker screening. Because of the abundant amino groups of acrylic acid (AA) component, the constructed hydrogel PhC barcodes with inverse opal structure could facilitate the loading of antibody probes for subsequent detection of tumor markers. By integrating tyramine deposition amplified ELISA on the barcode, the detection signal of tumor markers has been enhanced. Based on these features, it is demonstrated that the hydrogel PhC barcodes with tyramine deposition amplified ELISA could realize highly sensitive and multiplexed detection of HCC-related biomarkers. It was found that this method is flexible, sensitive and accurate, suitable for multivariate analysis of low abundance tumor markers and future cancer diagnosis. These features make the newly developed PhC barcodes an innovation platform, which possesses tremendous potential for practical application of low abundance targets.
研究领域:
生物医学工程
·
癌症研究
·
分子诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Toxins
·
2025
·Vol. 17
·期 9
·426
·
IF 4
Jie Wang,Wangzhuo Fu,Xuezhen Ma,Lin Chen,Weitao Song,Sumei Ling,Hongyun Qian,Shihua Wang,Zhenhong Zhuang
Due to the severe hazard of aflatoxins (AFs) to humans, it is of great significance to detect the key aflatoxins, aflatoxin B1(AFB1) and aflatoxin G1(AFG1), in food and feed in simple, rapid, and semi-quantitative ways. The hybridoma clone 3A1 was prepared in this study, and anti-AFB1monoclonal antibody (mAb) with high specificity and affinity (9.38 × 108L/mol) from 3A1 was purified. The indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) demonstrated that the linear detection range for AFB1was 0.029–1.526 ng/mL with a limits of determination (LOD) of 0.023 ng/mL. A latex microsphere-based immunochromatographic test strip (LM-ICTS) was constructed based on 3A1, which showed that the strip could detect AFB1(LOD: lower than 1.79 ng/mL) and AFG1(LOD: lower than 8.08 ng/mL), and the linear detection ranges for AFB1and AFG1are 1.79–48.46 ng/mL and 8.08–107.40 ng/mL, respectively. The average recoveries of intra-assay and inter-assay for peanuts were (98.4 ± 4.7)% and (92.6 ± 7.6)%, and the average coefficient of variation (CVs) were 4.38% and 8.15%, respectively. For sunflower seeds, the intra-assay and inter-assay recoveries were (94.4 ± 7.2)% and (89.2 ± 4.3)%, and the average CVs were 6.6% and 4.9%, respectively. In summary, the developed LM-ICTS exhibited excellent sensitivity and specificity, which provided a rapidly stable on-site detection choice for AFB1and AFG1to contaminated agricultural samples, including grain and feed.
研究领域:
免疫学
·
食品安全
·
分析化学
·
分子生物学
·
农产品质量检测
·
毒理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYST
·
2021
·Vol. 146
·期 22
·6831-6839
·
IF 4.616
Huihui Wu,Xinxin Xu,Liqiang Liu,Liguang Xu,Hua Kuang,Chuanlai Xu
In this study, a highly specific and sensitive monoclonal antibody (mAb) against quinclorac (Qui) was prepared. Based on the selected mAb, 2G3, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and an immunochromatographic strip assay were established for the detection of Qui in actual samples. The 50%-inhibitory concentration of mAb 2G3 against Qui was 48.763 ng mL−1. No cross-reaction with other quinolines indicated that mAb 2G3 had high specificity. The recovery of the established ic-ELISA method was in the range of 85.6% to 98.9%. The cut-off value of Qui in cucumber and tomato by immunochromatographic strip was 200 ng g−1. The analysis results of ic-ELISA and immunochromatographic strip assay were consistent with the results of LC-MS/MS, which further proved that the established ic-ELISA and immunochromatographic strip assay could provide valuable tools for the rapid detection of Qui residues in cucumber and tomato samples.
研究领域:
农业化学
·
食品科学
·
分析化学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF IMMUNOLOGICAL METHODS
·
2024
·
IF 1.6
Fang Lu,Xiqun Wu,Fa Zhang,Jiaqiang Wu,Zhaodong Yuan,Baomin Wang,Guiyu Tan,Suqin Guo
Immunoassay relies on antibodies, but traditional antibodies such as monoclonal antibody (mAb) require animal immunization and complex procedures. Single-chain variable fragment (scFv) becomes a potential alternative to mAb with advantages of the low cost, rapid and easy prepared. In the present study, we prepared scFvs against dihydroartemisinin (DHA) based on E. coli and HEK293T cell expression system, named MBP-scFv and scFv-Fc, respectively. Their properties were compared with the parent mAb. The calculated affinity constants of mAb, MBP-scFv and scFv-Fc were 2.1 × 10 8 L mol −1 , 2.2 × 10 7 L mol −1 and 1.6 × 10 8 L mol −1 , respectively. The half inhibitory concentration (IC 50 ) of mAb, MBP-scFv and scFv-Fc were 1.16 ng mL −1 , 2.15 ng mL −1 and 6.57 ng mL −1 , respectively. Both the scFv showed less sensitive than the mAb based on the IC 50 . The cross-reactivities of MBP-scFv for artemisinin and artesunate exhibited similarities to the mAb, yet the cross-reactivities of scFv-Fc for these compounds exceeded those of the mAb significantly. The stability of the scFvs was ascertained to be maintained for over 5 days at room temperature, and for more than a month at both 4 °C and − 20 °C. After that, the indirect competitive enzyme-linked immunosorbent assays (icELISAs) based on the scFv from E. coli were used to detect the DHA content in eight drug samples, and the result was consistent with ultra-performance liquid chromatography simultaneously. Although scFv can be used for quantitative determination of drugs, but it still cannot completely replace mAb in immunoassay without evolution and modification.
研究领域:
免疫学
·
药学
·
分子生物学
✓ 已验证
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）