JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2014
·
IF 3.107
Lin Luo,Zhen-Lin Xu,Jin-Yi Yang,Zhi-Li Xiao,Yong-Jun Li,Ross C. Beier,Yuan-Ming Sun,Hong-Tao Lei,Hong Wang,Yu-Dong Shen
Novel haptens were designed and synthesized to prepare antibodies against free histamine, but none resulted in producing suitable antibodies for developing an enzyme-linked immunosorbent assay (ELISA). However, an antiserum was obtained having high specificity and affinity to p-nitrobenzoylated histamine (NPHA), which can be easily formed from reaction between histamine and p-nitrobenzoic acid N-hydroxysuccinimide ester (PNBA-OSu) under mild conditions. Based on rabbit polyclonal antibodies, a competitive indirect ELISA (ciELISA) for histamine determination in foods was developed. After ciELISA and derivatization optimization, the assay showed good sensitivity, with limits of detection of 1.8 mg/kg, 93.6 μg/L, and 93.6 μg/kg in fish, red wine, and yoghurt, respectively, with negligible cross-reactivity with related biogenic amines and amino acids. Average recovery of histamine in fortified food samples ranged from 80.9% to 110.1% with coefficients of variation below 16.3%. Good correlation between the ciELISA and liquid chromatography–tandem mass spectrometry was obtained for spiked food samples.
研究领域:
食品科学
·
免疫学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2019
·
IF 6.393
Hongye Yan,Lei Jiao,Hengjia Wang,Weiqing Xu,Yu Wu,Wenling Gu,Dan Du,Yuehe Lin,Chengzhou Zhu
Enzyme-linked immunosorbent assay (ELISA) has been widely used in the area of biological analysis, while there are still some challenges getting in the way to ensure both excellent sensitivity and reliable accuracy. To overcome these obstacles, we simulated the “smart” sense-and-treat carriers in a drug delivery system to fabricate an improved ELISA, employing zeolitic imidazolate framework-8 (ZIF-8) as the carrier to deliver the tracer agent carbon dots (CDs) and the “drug” thymolphthalein (TP), which was referred to as a “sense-and-treat” ELISA. In the “sense” part, the strong fluorescence intensity of CDs could be observed directly to achieve the sensitive detection of the target. In the “treat” part, after stimulation of alkaline solution, TP was released from ZIF-8 carriers and generated a color change with an obvious absorption, which was beneficial for increasing the sensitivity of this ELISA due to the high loading of the TP. Finally, we took carcinoembryonic antigen (CEA) as a model and performed dual-modal detection using this improved “sense-and-treat” ELISA to accomplish quantitative determinations sensitively and accurately. In addition, the results indicated by the two parts kept good concordance, which could demonstrate the reliable accuracy of this ELISA.
研究领域:
生物分析
·
医学诊断
✓ 已验证
产品关联
DOI
高影响因子
Foods
·
2021
·Vol. 10
·期 10
·2398
·
IF 4.35
Xiya Zhang,Mingyue Ding,Chensi Zhang,Yexuan Mao,Youyi Wang,Peipei Li,Haiyang Jiang,Zhanhui Wang,Xuezhi Yu
The consumption of shellfish contaminated with brevetoxins, a family of ladder-frame polyether toxins formed during blooms of the marine dinoflagellateKarenia brevis, can cause neurotoxic poisoning, leading to gastroenteritis and neurotoxic effects. To rapidly monitor brevetoxin levels in oysters, we generated a broad-spectrum antibody against brevetoxin 2 (PbTx-2), 1 (PbTx-1), and 3 (PbTx-3) and developed a rapid indirect competitive enzyme-linked immunosorbent assay (icELISA). PbTx-2 was reacted with carboxymethoxylamine hemihydrochloride (CMO) to generate a PbTx-2-CMO hapten and reacted with succinic anhydride (HS) to generate the PbTx-2-HS hapten. These haptens were conjugated to keyhole limpet hemocyanin (KLH) and bovine serum albumin (BSA) to prepare immunogen and coating antigen reagents, respectively, using the active ester method. After immunization and cell fusion, a broad-spectrum monoclonal antibody (mAb) termed mAb 1D3 was prepared. The 50% inhibitory concentration (IC50) values of the icELISA for PbTx-2, PbTx-1, and PbTx-3 were 60.71, 52.61, and 51.83 μg/kg, respectively. Based on the broad-spectrum mAb 1D3, an icELISA was developed to determine brevetoxin levels. Using this approach, the limit of detection (LOD) for brevetoxin was 124.22 μg/kg and recoveries ranged between 89.08% and 115.00%, with a coefficient of variation below 4.25% in oyster samples. These results suggest that our icELISA is a useful tool for the rapid monitoring of brevetoxins in oyster samples.Keywords:brevetoxins;monoclonal antibody;enzyme-linked immunosorbent assay
研究领域:
海洋生物学
·
免疫学
·
食品安全
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF FOOD QUALITY
·
2020
·
IF 1.763
Shengyun Li,Yaowu Yuan,Chenchen Yu,Hao Gao,Jianxin Tan,Yiling Tian
A method for the quantitative determination of ganoderic acid A was constructed using the principle of indirect competitive enzyme-linked immunosorbent assay (ELISA), and this method was used to determine the ganoderic A contents of Ganoderma lucidum samples in the market. The conjugate of ganoderic acid A and bovine serum albumin was used for four rounds of immunization on test rabbits to obtain rabbit antiganoderic acid A antibody IgG. The enzyme-labeled plate was coated with the conjugate of ganoderic acid A and ovalbumin. The first stage reaction in the indirect competitive ELISA was that the conjugate of ganoderic acid A in the sample competed with the conjugate coated on the enzyme-labeled plate to bind rabbit antibodies. The second stage reaction was the combination of goat anti-rabbit IgG–horseradish peroxidase and rabbit antiganoderic acid A antibody IgG. The results of the determination of ganoderic acid A standard by this method showed that the coefficient of variation of repeated wells in the group was <5%, the detection limit of ganoderic acid A was 0.6 μg/L, and ganoderic acid A had a substantial dose-response relationship in the content range of 0.9–72.9 μg/L (R2 = 0.994). This method was used to measure the ganoderic A content of 12 varieties of G. lucidum in the market and showed the obvious differences in the ganoderic acid A contents of the different varieties. This method is simple, fast, and of great importance to the quality control of Ganoderma products.
研究领域:
分析化学
·
药理学
·
真菌学
✓ 已验证
产品关联
DOI
高影响因子
Materials
·
2023
·Vol. 16
·期 13
·4641
·
IF 3.4
Fei Fang,Hai-Yang Zhao,Rui Wang,Qi Chen,Qiong-Yan Wang,Qing-Hua Zhang
The abnormal increase in low-density lipoprotein (LDL) in human blood is a main independent risk factor for the pathogenesis of atherosclerosis, whereas a reduced LDL level effectively lowers morbidity. It is important to develop LDL adsorption materials with high efficiency and selectivity, as well as to simplify their fabrication processes. In this paper, polysulfone (PSF), sulfonated polysulfone (SPSF), and sulfonated polysulfone/dextran (SPSF/GLU) membranes were successfully fabricated for LDL adsorption using a solution casting technique. Attenuated total reflectance Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy measurements confirmed the success of the preparation. The water contact angle decreased from 89.7 ± 3.4° (PSF) to 76.4 ± 3.2° (SPSF) and to 71.2 ± 1.9° (SPSF/GLU), respectively. BSA adsorption testing showed that the SPSF/GLU with surface enrichment of sulfonate groups and glycosyl groups possessed higher resistance to protein solution. The adsorption and desorption behaviors of the studied samples in single-protein or binary-protein solutions were systematically investigated by enzyme-linked immunosorbent assay (ELISA), The results showed that SPSF/GLU, which had excellent resistance to protein adsorption, possessed a similar adsorption capacity to that of PSF. SPSF membrane exhibited excellent selective affinity for LDL in single and binary protein solutions, suggesting potential applications in LDL removal.Keywords:atherosclerosis;polysulfone;low-density lipoprotein;enzyme-linked immunosorbent assay
研究领域:
生物医学工程
·
材料科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL BIOCHEMISTRY
·
2021
·
IF 3.365
Yuanyuan Ding,Xiaoqian Li,Qingpeng Gao,Xinyan Dong,Liyun Kong,Shengli Han,Tao Zhang,Langchong He
Mas-related G-protein-coupled receptor X2 (MRGPRX2) has recently been reported to be associated with anaphylaxis. Detection of MRGPRX2 levels in human peripheral blood might serve as a powerful tool for predicting the predisposition of patients to anaphylactic reactions. For rapid measurement of MRGPRX2, we established a paper-based double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) using mouse monoclonal antibody and horseradish peroxidase (HRP)-labelled rabbit polyclonal antibody as capture antibody and detection antibody, respectively. We avoided chemical functionalization of the cellulose paper by introducing bovine serum albumin (BSA) to provide COOH and NH 2 groups for covalent immobilization of the capture antibody. Through amide condensation , a two-layer immobilization strategy was applied with BSA-BSA and BSA-capture antibody networks as the first and second layers, respectively. This strategy improved the quantity, activity and stability of the immobilized antibody . We then established a paper-based ELISA to detect MRGPRX2 in human peripheral blood. Our method is less laborious, easier to implement, and more cost-effective than conventional ELISA, while offering similar sensitivity, specificity, and accuracy. Therefore, it could serve as an innovative clinical point-of-care diagnostic tool, especially in areas that lack advanced clinical equipment.
研究领域:
医学
·
免疫学
·
生物化学
✓ 已验证
DOI
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
·
2011
·
IF 0.633
Fei Lin,Shanshan Song,Liqiang Liu,Hua Kuang,Libing Wang,Chuanlai Xu
An enzyme-linked immunosorbent assay method was developed here for benzophenone (BZP) detection in package paper samples based on polyclonal antibody. 4-Aminobenzophenone (ABZP) was used as hapten and conjugated with bovine serum albumin for antibody production. Cross-reactivity results showed that the antibody can be recognised BZP, ABZP and Michler's ketone very well (IC50 value was 31.2±7.8 ng/mL, 35.9±8.3 ng/mL and 43.7±8.2 ng/mL, respectively). After optimisation of the effects from pH, ionic strength and organic solvents on immunoassay, we established a rapid method and used for BZP detection in fortified package paper samples with good recovery data (98.5 and 99.7%) at 100 and 600 ng/g spiked level.)
研究领域:
环境科学
·
分析化学
✓ 已验证
DOI
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2019
·
IF 2.813
Jiawen Li,Dongqiang Lin,Shanjing Yao,N.K.H. Slater,Qilei Zhang
Traditional chromatographic techniques used in downstream processes of biomolecule manufacturing are often time-consuming and expensive. In this study, a cost-effective microporous micro-capillary film (MMCF) composed of ethylenevinyl alcohol (EVOH) was evaluated for its potential application in immunoadsorption with high process efficiency. A peptide ligand Ac-Phe-Tyr-His-Glu (Ac-FYHE) was immobilized on the inner surface of MMCF for selective binding of human immunoglobulin (hIgG). The porous structure and chemical properties of the prepared MMCF were confirmed by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). hIgG (2 mg/ml) adsorption studies demonstrated that the binding process followed a Langmuir isotherm with equilibrium adsorption capacities of 9.31 and 3.47 mg/ml adsorbent under static and dynamic conditions, respectively. Moreover, the membrane showed good flowrate tolerance when studied under flowrates of 0.5 ml/min to 10 ml/min. hIgG purity was 88.2% when obtained from an hIgG (2 mg/ml) and HSA (8 mg/ml) mixture and the purity remained over 80.0% when hIgG concentrations increased in the mixtures. Moreover, purity of 82.3% was achieved when removing hIgG directly from human serum. The MMCF-Ac-FYHE affinity column is expected to selectively remove hIgG from blood for the treatment of autoimmune diseases with high efficiency and cost effectiveness.
研究领域:
生物工程
·
医学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
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
MOLECULES
·
2019
·Vol. 24
·期 11
·2119
·
IF 3.06
Lijun Zhang,Berlin Zang,Chundong Huang,Jun Ren,Lingyun Jia
Dialysis-related amyloidosis (DRA), which has been widely recognized to be associated with the accumulation of β2-microglobulin (β2-m) in blood, is one of the most common complications in patients receiving long-term dialysis treatment. The most significant side-effect of existing hemodialysis sorbents for the removal of β2-m from blood is the loss of vital proteins due to non-specific adsorptions. Although the traditional antibodies have the capability to specifically remove β2-m from blood, high cost limits their applications in clinics. Single domain antibodies derived from the Camelidae species serve as a superior choice in the preparation of immunoadsorbents due to their small size, high stability, amenability, simplicity of expression in microbes, and high affinity to recognize and interact with β2-m. In this study, we modified the anti-β2-m VHH by the formylglycine-generating enzyme (FGE), and then directly immobilized the aldehyde-modified VHH to the amino-activated beads. Notably, the fabrication is cost- and time-effective, since all the preparation steps were performed in the crude cell extract without rigorous purification. The accordingly prepared immunoadsorbent with VHHs as ligands exhibited the high capacity of β2-m (0.75 mg/mL). In conclusion, the VHH antibodies were successfully used as affinity ligands in the preparation of novel immunoadsorbents by the site-specific immobilization, and effectively adsorbed β2-m from blood, therefore opening a new avenue for efficient hemodialysis.Keywords:β2-microglobulin;VHH;formylglycine-generating enzyme;hemodialysis
研究领域:
生物医学工程
·
分子生物学
·
透析治疗
✓ 已验证
DOI
PubMed
高影响因子
ARCHAEOMETRY
·
2019
·Vol. 61
·期 4
·921-932
·
IF 1.64
H. L. Zheng,C. Ma,X. J. Dai,Y. Zhou
Identifying the traces and residues of unearthed silk fibres is crucial for further analysis and study. In this study, an enzyme-linked immunosorbent assay (ELISA) based on the monoclonal antibodies specific to Bombyx mori fibroin was developed in order to for detect ancient silk. Five monoclonal antibodies were successfully prepared based on fibroin hydrolysate, and four had a high specificity for mulberry silk. Furthermore, they could be used to discern B. mori fibre from tussah fibre and eri fibre. The ELISA standard curve of the most sensitive monoclonal antibody, anti-Silk Fibroin (SF)-3, was y = 0.022 x + 0.177 ( R 2 = 0.998), and the detection limit was 4.65 ng/ml. Using this antibody, we successfully identified five ancient mulberry silk samples from different time periods. The results suggest that ELISA assays based on monoclonal antibodies can be used as an immunological testing method for unearthed silk fibre.
研究领域:
考古学
·
免疫学
·
材料科学
✓ 已验证
DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2021
·
IF 10.618
Lei Chen,Yan Li,Luyang Miao,Xiaolong Pang,Tao Li,Yongjun Qian,He Li
In the field of precision medicine, the anticipated features of ideal drug delivery systems (DDS) have high drug loading capacity and effective stimuli-triggered mechanism, which are fitting well with the expected merits of signal labels for enhanced enzyme-linked immunosorbent assay (ELISA). Inspired by this, poly (diallyldimethylammonium chloride)-capped curcumin nanoparticles (PDDA@CUR NPs) with high loading capacity were synthesized as signal labels and further applied to dual-model colorimetric and fluorescence ELISA for the detection of C-reactive protein (CRP). Curcumin (CUR) was elaborately selected as report molecule similar to the roles of drugs in DDS, which dispersed in neutral water exhibits a negligible fluorescence response due to the aggregation of CUR molecules induced quenching effect, stimulated by basic water (BW, pH 12.36), the allochroic effect from colorless to orange occurred and fluorescence restored because of the keto–enol tautomerism in the molecular structure of CUR, just like lighting-up (from signal “OFF” to signal “ON”), yielded a dual-model colorimetric and fluorescent signal readout. PDDA, as a polycationic electrolyte, provided a biological platform that is capable of interacting with CRP label antibodies by virtue of its positive centers. The results show that “lighting-up” CUR NPs-based dual-modal colorimetric and fluorescent ELISA for CRP detection has the merits of easy-to-use, good enough sensitivity and reliability. And more importantly, it brings innovative ideas for the precise identification and quantification of protein biomarkers.
研究领域:
生物医学工程
·
诊断学
✓ 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）