FOOD CHEMISTRY
·
2020
·
IF 6.306
Min Jia,Junyi Sha,Zhihua Li,Wenjing Wang,Hongyan Zhang
The widespread exposure of bisphenol A (BPA) presents a significant risk to human health. A rapid, ultra-sensitive and label-free colorimetric aptasensor using high affinity truncated aptamers was developed for BPA detection. Truncated 38-mer and 12-mer aptamers specific for BPA were obtained through rationally truncation from 63-mer BPA aptamer. The dissociation constants ( K d ) of 38-mer and 12-mer aptamers were determined to be 13.17 nM and 27.05 nM. Then, truncated aptamers were used in label-free colorimetric detection assays based on gold nanoparticles (AuNPs). The limit of detections of aptasensors using 38-mer and 12-mer aptamers were 7.60 pM and 14.41 pM, which were 265-fold and 140-fold lower than that of the aptasensor using 63-mer aptamer, respectively. The recovery rates in milk, orange juice and mineralized water samples were 93.88% to 107.30%. Therefore, the developed BPA colorimetric aptasensor using truncated aptamers has great application prospects in food safety control and environmental monitoring.
研究领域:
分析化学
·
环境科学
·
食品安全
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产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2019
·
IF 5.399
Chan-Yuan Yao,Zhen-Lin Xu,Hong Wang,Fan Zhu,Lin Luo,Jin-Yi Yang,Yuan-Ming Sun,Hong-Tao Lei,Yuan-Xin Tian,Yu-Dong Shen
Alternariol (AOH), a secondary metabolites of the genus Alternaria , is a genotoxic mycotoxin. Because the previous “Mannich antigen” for AOH readily underwent unstable induction, a new AOH hapten was designed for inducing a high-quality antibody against AOH using a conjugate of the AOH carboxymethyl derivative with a carrier protein as the immunogen. A competitive indirect chemiluminescence enzyme immunoassay (ciCLEIA) for AOH was optimized and showed good sensitivity (limit of detection 0.068 ng/mL), a linear range (0.11–1.23 ng/mL), and negligible cross-reactivity with analogues. Average rates of recovery from spiked samples (juice and cereal) were between 72.7% and 115.8%, with coefficients of variation <14.3%. Results from ciCLEIA correlated well with those of high-performance liquid chromatography coupled with tandem mass spectrometry, meaning the proposed method might be an effective screening tool for detection of AOH in food samples.
研究领域:
食品科学
·
免疫学
·
毒理学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF SEPARATION SCIENCE
·
2016
·Vol. 39
·期 16
·3254-3259
·
IF 2.741
Zhengzhong Lin,Dan Wang,Aihong Peng,Zhiyong Huang,Yuhui Lin
A selective sample cleanup method using molecularly imprinted polymers was developed for the separation of domoic acid (a shellfish toxin) from shellfish samples. The molecularly imprinted polymers for domoic acid was prepared by emulsion polymerization using 1,3,5-pentanetricarboxylic acid as the template molecule, 4-vinyl pyridine as the functional monomer, ethylene glycol dimethacrylate as the crosslinker, and Span80/Tween-80 (1:1 v/v) as the composite emulsifiers. The molecularly imprinted polymer showed high affinity to domoic acid with a dissociation constant of 13.5 μg/mL and apparent maximum adsorption capacity of 1249 μg/g. They were used as a selective sorbent for the detection of domoic acid from seafood samples coupled with high-performance liquid chromatography. The detection limit of 0.17 μg/g was lower than the maximum level permitted by several authorities. The mean recoveries of domoic acid from clam samples were 93.0–98.7%. It was demonstrated that the proposed method could be applied to the determination of domoic acid from shellfish samples.
研究领域:
分析化学
·
食品安全
·
海洋生物学
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产品关联
DOI
PubMed
高影响因子
Journal of the American Chemical Society
·
2022
·
IF 16.383
Guohua Xu,Jiajing Zhao,Hao Yu,Chen Wang,Yangyu Huang,Qiang Zhao,Xin Zhou,Conggang Li,Maili Liu
A 36-mer guanine (G)-rich DNA aptamer (OBA36) is able to distinguish one atomic difference between ochratoxin analogues A (OTA) and B (OTB), showing prominent recognition specificity and affinity among hundreds of aptamers for small molecules. Why OBA36 has >100-fold higher binding affinity to OTA than OTB remains a long-standing question due to the lack of high-resolution structure. Here we report the solution NMR structure of the aptamer–OTA complex. It was found that OTA binding induces the aptamer to fold into a well-defined unique duplex–quadruplex structural scaffold stabilized by Mg2+ and Na+ ions. OTA does not directly interact with the G-quadruplex, but specifically binds at the junction between the double helix and G-quadruplex through π–π stacking, halogen bonding (X-bond), and hydrophobic interaction. OTB has the same binding site as OTA but lacks the X-bond. The strong X-bond formed between the chlorine atom of OTA and the aromatic ring of C5 is the key to discriminating the strong binding toward OTA. The present research contributes to a deeper insight of aptamer molecular recognition, reveals structural basis of the high-affinity binding of aptamers, and provides a foundation for further aptamer engineering and applications.
研究领域:
生物化学
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分子生物学
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结构生物学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2025
·Vol. 26
·期 5
·2119
·
IF 4.9
Chengwei Wang,Chunjie Wu,Linjiang Song
Saiga antelope horn (SAH), a rare traditional Chinese medicine, exhibits activities of anti-feverish convulsions and anti-inflammation, whereas its underlying mechanism and specific pharmacological components are still unclear. In the present study, transient receptor potential ankyrin 1 (TRPA1), a major transient receptor potential cation channel was used as a target protein to identified TRPA1 high-affinity peptides (THPs) from SAH digests. Firstly, the SAH was digested under in vitro gastrointestinal conditions. With the method of affinity ultrafiltration and liquid chromatography–mass spectrometry (AUF-LC/MS), about 200 peptides that have a high-affinity interaction with the TRPA1 protein were screened from SAH digests. Subsequently, bioactivity databases and molecular docking were further exploited to identified three THPs, including RCWPDCR, FGFDGDF, and WFCEGSF. Furthermore, RIN-14B cells, characterized by the high expression of TRPA1 on cell surfaces, were used as the cell model to investigate the biological effect of THPs. Immunofluorescence and ELISA were conducted and showed that THPs can increase the intracellular Ca2+concentration and serotonin (5-HT) secretion in RIN-14B cells by activating TRPA1, which is evidenced by impaired upregulation of intracellular Ca2+levels and 5-HT secretion after pretreatment with the TRPA1 inhibitor (HC-030031). Moreover, an analysis of Western blots displayed that THPs up-regulated the expression levels of the 5-HT synthesis rate-limiting enzyme (TPH1) and 5-hydroxytryptophan decarboxylase (Ddc), while serotonin reuptake transporter (SERT) levels were down-regulated, suggesting that THPs enhance 5-HT secretion by regulating the 5-HT synthesis pathway. In summary, our findings demonstrate that THPs, which were identified from SAH digest via TRPA1-targeted affinity panning, exhibited the activation of the TRPA1 channel and enhanced 5-HT release in RIN-14B cells.
研究领域:
药理学
·
分子生物学
·
传统中药
·
细胞生物学
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产品关联
DOI
PubMed
高影响因子
Frontiers in Immunology
·
2019
·
IF 4.716
Wu Liting,Fu Shengli,Yin Xiaoxue,Guo Zheng,Wang Anli,Ye Jianmin
The recent discovery of long-lived plasma cells (LLPCs) in mammals, which provide a constant expression of specific high-affinity antibodies that mediate humoral memory, has caused a dramatic paradigm shift in the study of immunity and vaccine development. In teleost fish, there are few studies regarding the association between LLPCs and antibody production, and the affinity of the antibodies secreted by the LLPCs is poorly understood. In the present study, channel catfish (Ictalurus punctatus) were immunized with trinitrophenylated-keyhole limpet hemocyanin (TNP-KLH) to examine TNP-specific antibody titers, affinities, antibody-secreting cell (ASC) dynamic changes, and especially the affinity of secreted antibodies by LLPCs post-immunization. We demonstrated that TNP-specific LLPCs were generated starting at week 4 post-immunization, achieved a maximal number at week 8, and maintained a comparable level throughout the 18-week post-immunization period, which was correlated with the dynamics of serum antibody titers and affinity maturation in the response. The LLPCs appeared to mostly reside within, or migrate to, the anterior kidney (bone marrow-like tissue in mammals), but a small portion was also located in the spleen and peripheral blood. The antibodies produced by the LLPCs possessed high affinities, indicating that the generation and development of LLPCs were driven by affinity selection in teleosts. Collectively, the results of this study provide insights toward the evolutionary understanding of the affinity-dependent mechanism of LLPC generation and development.
研究领域:
免疫学
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兽医学
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水生生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
RSC Advances
·
2019
·Vol. 9
·期 11
·6328-6334
·
IF 3.049
Qiong-Lin Wang,Hui-Fang Cui,Jiang-Feng Du,Qi-Yan Lv,Xiaojie Song
DNA aptamers against carcinoembryonic antigen (CEA) have been identified through the systematic evolution of ligands by exponential enrichment (SELEX) technique, but their affinity needs to be improved. In this study, an in silico approach was firstly used to screen the mutation sequences of a reported DNA aptamer (the parent aptamer, denoted as P) against CEA. The affinities of several high-score DNA mutants were determined by the biolayer interferometry technique. Finally, the newly obtained aptamers were verified in an aptasensor application. For the in silico approach, Mfold and RNA Composer were combined to generate the 3D RNA structures of the DNA mutants. The RNA structures were then modified to 3D DNA structures with the Write program. The docking model and binding ability of the 3D DNA structures with CEA were simulated and predicted with the ZDOCK program. Two mutation sequences (P-ATG and GAC-P) exhibited significantly higher ZDOCK scores than P. The dissociation constant of P-ATG and GAC-P to CEA was determined to be 4.62 and 3.93 nM respectively, obviously superior to that of P (6.95 nM). The detection limit of the P-ATG and GAC-P based aptasensors was 1.5 and 1.2 ng mL−1, respectively, markedly better than that based on P (3.4 ng mL−1). The consistency between the in silico and the experimental results indicates that the developed in silico post-SELEX screening approach is feasible for improving DNA aptamers. The P-ATG and GAC-P aptamers found in this study could be used for future CEA aptasensor design and fabrication, promisingly applicable for highly sensitive CEA detection and early cancer diagnosis.
研究领域:
生物信息学
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分子生物学
·
癌症诊断
·
纳米技术
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2019
·
IF 6.35
Wei Li,Qi Zhang,Yijia Wang,Yanyan Ma,Zhanchen Guo,Zhen Liu
Molecularly imprinted polymers (MIPs) and aptamers, as effective mimics of antibodies, can overcome only some drawbacks of antibodies. Since they have their own advantages and disadvantages, the combination of MIPs with aptamers could be an ideal solution to produce hybrid alternatives with improved properties and desirable features. Although quite a few attempts have been made in this direction, a facile and controllable approach for the preparation of aptamer–MIP hybrids still remains lacking. Herein, we present a new approach for facile and controllable preparation of aptamer–MIP hybrids for high-specificity and high-affinity recognition toward proteins. An aptamer that can bind the glycoprotein alkaline phosphatase (ALP) with relative weak affinity and specificity was used as a ligand, and controllable oriented surface imprinting was carried out with an in-water self-polymerization system of dopamine. A thin layer of polydopamine was formed to cover the template to an appropriate thickness. After removing the template from the polymer, an aptamer–MIP hybrid with apparently improved affinity and specificity toward ALP was obtained, giving cross-reactivity of 3.2–5.6% and a dissociation constant of 1.5 nM. With this aptamer–MIP hybrid, a plasmonic immunosandwich assay (PISA) was developed. Reliable detection of ALP in human serum by the PISA was demonstrated.
研究领域:
生物材料
·
分析化学
·
生物技术
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2022
·
IF 16.744
Sijie Liu,Rui Shu,Jiaqi Ma,Leina Dou,Wentao Zhang,Shaochi Wang,Yanwei Ji,Yuechun Li,Jingke Xu,Daohong Zhang,Mingqiang Zhu,Yumei Song,Jianlong Wang
Lateral flow immunoassay (LFIA) is a paper-based point-of-care (POC) diagnostic tool for widespread applications. Nonetheless, the lack of multi-target monitor capability, high-affinity between signal tag and antibody, and multi-signal readout capability still remain challenges. Herein, a mussel-inspired Fe-based Tannic acid (TA) Nanozyme (FTAN) mediated dual-readout and dual-semiquantitative multiplex lateral flow immunoassay (MLFIA) was strategically fabricated by employing ractopamine (RAC) and clenbuterol (CLE) as proof-of-concept. Avail of the environment-friendly, cost-effective, darker original color, and dual-signal readout capacity, FTAN was innovatively designed by chelating renewable bioresource TA as coupling element and Fe as active center to manufacture a multi-functional signal tag for the first time. Benefiting from the preferential protein adsorption ability of TA-Fe, the mussel-inspired nanozyme achieves satisfactory coupling efficiency and stability with antibodies. Importantly, the whole detection range was divided into several monitoring sections owing to the defined multiple detection limits (originated from on-demand outputs of colorimetric and catalytic signal) and broadened detection range, which contribute to satisfies dual-semiquantitative rapid on-site detection and various limit standards of same target in different regions. This research may contribute to the rational design of high-affinity nanozyme-based signal tags in LFIAs, the exploration of more renewable bioresource-derived nanozymes, and the extension of their applications in relevant sectors.
研究领域:
分析化学
·
纳米技术
·
生物医学工程
·
即时诊断
·
生物化学
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DOI
高影响因子
JOURNAL OF SEPARATION SCIENCE
·
2024
·Vol. 47
·期 18
·e202400471
·
IF 2.8
Qian Qin,Xiaolong Liu,Xun Wang,Lina Zhou,Huihui Wan,Qingxin Yin,Di Chen
Recombinant proteins hold significant importance in numerous disciplines. As the demand for expressing and purifying these proteins grows, the scientific community is in dire need of a simple yet versatile methodology that can efficiently purify these proteins. Aptamers as synthetic nucleic acid–based ligands with high affinity have shown promise in this regard, as they can capture targets through molecular recognition. In this study, novel aptamer-functionalized polydopamine-coated magnetic graphene oxide nanocomposites were facilely prepared, achieving an impressive average aptamer coverage density (45 nmol/mg). These nanocomposites exhibited a uniform structure and robust magnetic responsiveness. The findings indicated that they possess several advantages, such as rapid adsorption, substantial capacity (171.4 mg/g), and excellent reusability. Notably, due to the inherent properties of nucleic acids, the immobilized aptamer-magnetic beads can be utilized repeatedly with high purification efficiency. Finally, the nanocomposites were further employed to purify His-tagged proteins from actual samples. Remarkably, they were able to selectively and efficiently isolate His-tagged retinoid X receptor alpha protein from complex Escherichia coli lysate. The purified His-tagged retinoid X receptor alpha protein was analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. This confirmed the efficacy of developed nanocomposites, reinforcing their vast potential for purification of His-tagged recombinant proteins.
研究领域:
生物技术
·
纳米技术
·
蛋白质工程
·
分析化学
·
材料科学
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产品关联
DOI
PubMed
高影响因子
DIGESTIVE DISEASES AND SCIENCES
·
2023
·Vol. 68
·期 6
·2414-2426
·
IF 3.1
Lin Mengjuan,Hu Guiying,Wang Zhengqiang,Yu Baoping,Tan Wei
Background The mechanism underlying irritable bowel syndrome (IBS), a common disease with hyperalgesia, remains elusive. The spinal cholinergic system is involved in pain modulation, but its role in IBS is unknown. Aims To determine whether high-affinity choline transporter 1 (CHT1, a major determinant of the cholinergic signaling capacity), is implicated in spinal modulation of stress-induced hyperalgesia. Methods A rat IBS model was established by water avoidance stress (WAS). Visceral sensations were detected by abdominal withdrawal reflex (AWR) and visceromotor response (VMR) to colorectal distension (CRD). Abdominal mechanical sensitivity was determined by von Frey filaments (VFFs) test. RT-PCR, Western blot, and immunostaining were performed for spinal CHT1 expression. Spinal acetylcholine (ACh) was measured by ELISA; the influence of spinal CHT1 on hyperalgesia were evaluated by intrathecal administration of MKC-231 (a choline uptake enhancer) and hemicholinium-3 (HC-3, a specific inhibitor of CHT1). Minocycline treatment was used to explore the role of spinal microglia in hyperalgesia. Results After 10 days of WAS, AWR scores and VMR magnitude to CRD, and the number of withdrawal events in VFF test were increased. Double-labeling showed that CHT1 in the dorsal horn was expressed in most of the neurons and almost all the microglia. The CHT1 expression and ACh levels in the spinal cord and the density of CHT1-positive cell in the spinal dorsal horn were enhanced in WAS-exposed rats. HC-3 enhanced pain responses in WAS rats; MKC-231 alleviated pain in WAS rats by upregulating CHT1 expression and increasing ACh production in the spinal cord. Furthermore, microglial activation in the spinal dorsal horn promoted the stress-induced hyperalgesia, and MKC-231 achieved analgesic effects by inhibiting the spinal microglial activation. Conclusions CHT1 exerts antinociceptive effects in spinal modulation of chronic stress-induced hyperalgesia by increasing ACh synthesis and suppressing microglial activation. MKC-231 has potential for treating disorders accompanied by hyperalgesia.
研究领域:
神经科学
·
疼痛研究
·
胃肠病学
·
药理学
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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）