JOURNAL OF MEMBRANE SCIENCE
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2017
·
IF 6.035
Jinxin Fan,Jianquan Luo,Yinhua Wan
Enzyme purification and subsequent immobilization usually require multiple steps and consume plenty of chemicals. Traditional dipping incubation operation is lengthy and the interior of porous carrier is not fully exploited due to the diffusion barrier . To solve these problems, one-step enzyme purification and immobilization was developed based on membrane chromatography concept. A membrane adsorber was prepared by polydopamine coating on a PVDF microfiltration membrane (or on a cheap and disposable cellulose filter paper) and subsequent polyethyleneimine grafting, without any solvent usage. Laccase was selectively immobilized on such membrane adsorber (enzyme activity: 39.9±4.4 U/mL; expressed activity: 19.6±3.0%) by directly capturing it from a crude fermentation broth without pH adjustment under flow-through mode. The immobilized laccase had a high purity of 92.2%, while the membrane permeability decreased due to the adsorption of laccase and impurities (“membrane fouling”). Thanks to the dominant convective transport, the enzyme loading was high and much faster than that by dipping incubation (5 min vs. 80 min). Moreover, enzyme reloading on membrane adsorber was easily realized by reversible desorption-adsorption process. The constructed biocatalytic membrane exhibited a commendable reusability for bisphenol A (BPA) removal only driven by its gravity.
研究领域:
生物化学
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环境科学
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材料科学
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产品关联
DOI
高影响因子
COLLOIDS AND SURFACES B-BIOINTERFACES
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2013
·
IF 3.554
Cang Wang,Yan Fan,Meng-Xin Hu,Wei Xu,Jian Wu,Peng-Fei Ren,Zhi-Kang Xu
Glycosylated membrane, as one of the most important affinity membranes, permits affinity separation/purification of proteins based on carbohydrate–protein interactions. It is an important scientific issue to screen facile method for fabricating the glycosylated membrane surface with high glycosyl density. Such a surface can be fabricated by the direct covalent immobilization of carbohydrate ligands on the surfaces of microporous polypropylene membrane (MPPM). First, alkyne-functionalized membrane surface was fabricated by plasma pretreatment combined with UV-induced graft polymerization of 3-(trimethylsilyl) propargyl methacrylate. Then, the glycosylated membrane surface was directly fabricated with the thiol–yne click reaction to ensure rapid process, improved efficiency, and high glycosyl density. Chemical and physical properties of the membrane surface were characterized by ATR/FT-IR, XPS, FESEM and water contact angle measurement. Static lectin adsorption indicates that the glycosylated membrane can specifically adsorb lectin concanavalin A (Con A) other than peanut agglutinin (PNA). Break through curves from dynamic Con A adsorption show the membrane has unique properties such as strong specificity, high adsorption capacity, and reversible binding capability. We suggest that the prepared glycosylated membrane is of great potentials in affinity membrane chromatography for rapid and high-resolution separation/purification of lectins.
研究领域:
生物化学
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材料科学
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分离技术
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DOI
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2025
·
IF 3.1
Liju Yu,Jie Gao,Tingting Deng,Chang Gao,Jiaxing Wang,Nan Wang
Heart failure is a global epidemic characterized by high mortality and morbidity rates. β1-Adrenoceptors (ADRβ1) antagonists are the main drugs used in the treatment of HF. A simple and convenient method for screening ADRβ1 antagonists is necessary. Nebivolol has four chiral centers, and its byproducts, which are not easy to avoid, may interact with ADRβ1. Cell membrane chromatography based on SNAP-tag technology provides a comprehensive solution. In this study, an ADRβ1 target cell membrane chromatography model based on SNAP-tag technology was established, and the selectivity, specificity, repeatability, and column life of the system were validated. Under optimized conditions, this model was used to screen for active components targeting ADRβ1 in nebivolol and its byproducts. The bioactivity of nebivolol and its byproducts was detected by calcium influx. The results showed that the ADRβ1 SNAP-tag/CMC model was successfully established and could be used to screen active drugs targeting ADRβ1. Among nebivolol and its byproducts, D-nebivolol (SRRR-enantiomer) had the best affinity for ADRβ1. Three byproducts of nebivolol also exhibited retention characteristics on ADRβ1. In vitro calcium influx experiments verified the interaction of these components with ADRβ1, showing good inhibitory effects on ADRβ1. In conclusion, the ADRβ1 SNAP-tag/CMC model developed in this study can be used for the comprehensive screening of active components targeting ADRβ1 in complex samples.
研究领域:
药理学
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药物筛选
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分子生物学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2024
·
IF 3.8
Li Chenjia,Liao Qi,Wang Rui,Zhang Xinping,Ma Mengyang,Liu Yonghong,Xiao Lei,Jiao Ying,Wang Nan
The scourge of drug addiction and abuse poses a significant challenge to society. Opioid drugs acting on μ-opioid receptor (OPRM1) make it one of the pivotal targets for drug addiction. In the past decade, sewage analysis has become a prevalent method of drug monitoring. However, traditional methods of detecting drugs in sewage are cumbersome, and rapid detection methods are relatively lacking. To address this, an innovative OPRM1-SNAP-tag/CMC method to directly identify drug components in sewage was established. Cell membrane chromatography (CMC) is an affinity chromatography technique which effectively detects receptor affinity substances. Cells constructed with high expression of specific receptor could be used to screen for compounds acting on the receptor. CMC based on OPRM1 provides a potentially convenient and effective tool for the detection of targeted drug components in sewage. In this study, the selectivity, reproducibility, column lifetime, and carryover of the CMC column had been assessed. Initially, we eluted the collected domestic sewage with methanol and acetonitrile, and the retention peaks were observed on the CMC system. Subsequently, without any preliminary sample preparation, we directly injected filtered samples of suspicious sewage into the OPRM1-SNAP-tag/CMC system, where we observed retention peaks as well. The retained components were further identified as morphine by using UPLC-MS/MS. In conclusion, the OPRM1-SNAP-tag/CMC method stands out as a reliable and robust model for the detection of drug components in sewage. It provides a valuable analytical tool for frontline drug control efforts, enhancing our capacity to monitor and mitigate the impact of drug abuse on society. Graphical Abstract
研究领域:
生物化学
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药物分析
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环境科学
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
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2017
·
IF 6.216
Jinxin Fan,Jianquan Luo,Yinhua Wan
Biocatalytic membranes are promising to remove micro-pollutants in aqueous environment due to their mild and green operation condition. However, more efforts need to be devoted to improving their removal efficiency and stability. In this study, metal chelating affinity membrane chromatography (MCAMC) was used to construct a biocatalytic membrane by selectively capturing laccase from a crude fermentation broth. Metal ions had a significant effect on the activity of the immobilized laccase and copper ion was the best choice. A pH of 4.5 was selected for laccase adsorption and its loading seemed the same under flow rates from 0.5 to 10 mL min −1 thanks to the inherent convective transport of membrane chromatography. The pH value and salt concentration in the storage buffer had an obvious effect on the stability of the immobilized laccase, and the prepared biocatalytic membrane retained 87% of initial activity after 20 days storage. When applying such membrane to micro-pollutant removal (taking bisphenol A (BPA) as an example), a high BPA removal efficiency (99.3%) could be obtained. The biocatalytic membranes could be operated at a high flux of 50 L m −2 h −1 without recycling the permeate into the feed, and its throughput and BPA removal rate were superior to the most results in the literature. However, BPA removal decline (from 99.6% to 56.6% after five cycles) occurred during the successive water treatment due to the membrane fouling caused by BPA polymerization products. Membrane regeneration could be achieved by simple elution-cleaning-reloading, and the laccase activity and BPA removal were fully recovered.
研究领域:
环境生物学
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生物工程
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DOI
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
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2020
·
IF 3.004
Fen Wei,Xiaoshuang He,Ke Xu,Sicen Wang
Cell membrane chromatography (CMC) is effective and widely used in drug screening, especially for the analysis of complex matrixes. However, it is time-consuming and costly given that cells or animals are employed for activity confirmation, which leads to a large amount of waste being produced if the result is negative. Stepwise frontal analysis is employed to saturate the affinity stationary phase, by using a series of low- to high-concentration solutions which resultantly form a staircase pattern. In doing so, the waste of samples, caused by the balancing process, can be avoided. In this study, stepwise frontal analysis coupled with a CMC system was performed for screening and characterizing the affinity of an active compound from wuweizi. Schizandrin A was screened and identified by α 1A AR /CMC coupled with UHPLC-MS/MS. By comparing the values obtained with those related to the equilibrium dissociation constant (K d ) calculated by zonal elution, the accuracy of the stepwise frontal analysis was verified. Subsequently, the type of affinity force between Schizandrin A and α 1A AR was studied by thermodynamic parameters. Moreover, schizandrin A showed an antagonistic effect on phenylephrine-induced contractions, which relax prostate muscle strips in a non-competitive antagonism manner. It has already suggested that the active compound, schizandrin A, could be used as a lead compound for the treatment of benign prostate hyperplasia (BPH) and should be further studied. Thus, the findings of this study are significant given that they could result in an online screening and affinity analysis method being utilized for the discovery of medicinal compounds as well as clarify the interaction characteristics between a drug and a receptor.
研究领域:
药物化学
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药理学
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中药学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2021
·Vol. 413
·期 7
·1917-1927
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IF 4.157
Zhang Liyang,Yi Xinyao,Wang Saisai,Liang Peida,Zhou Huaxin,Fu Jia,Jia Qianqian,Gao Jiapan,Lv Yanni,Han Shengli
A novel stability-enhanced graphene quantum dot (GQD)-decorated epidermal growth factor receptor (EGFR) cell membrane chromatography was constructed to study the potential application of GQDs in bioaffinity chromatography, and to screen active components acting on EGFR from traditional Chinese medicine (TCM). The carboxyl groups on the surface of GQDs reacted with the amino groups of the amino-silica gel ( SiO 2 -NH 2 ) to form a covalent bond, thereby preparing the GQD-decorated silica gel ( SiO 2 -GQDs ). The EGFR cell membrane was further immobilized on the SiO 2 -GQDs through the same covalent binding method to obtain the GQD-decorated cell membrane stationary phase ( SiO 2 -GQDs-CMSP ). In this way, the cell membrane was firmly immobilized on the decorated silica carrier. The life span and stability of the GQD-decorated cell membrane chromatographic ( SiO 2 -GQDs-CMC ) column were both enhanced, and the optimal immobilization conditions of the EGFR cell membrane were also determined. This model was then verified by establishing a SiO 2 -GQDs-CMC online liquid chromatography-ion trap-time-of-flight (LC-IT-TOF) system to screen possible active components in Peucedanum praeruptorum Dunn . As a result, praeruptorin B (Pra-B) was screened out, and its inhibitory effect against EGFR cell growth was evaluated by the cell counting kit-8 (CCK-8) assay. Molecular docking assay was also conducted to further estimate the interaction between Pra-B and EGFR. Overall, this research indicated that GQDs may be a promising nanomaterial to be used in prolonging the life span of the CMC column, and Pra-B could be a potential EGFR inhibitor so as to treat cancer. Graphical abstract
研究领域:
分析化学
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药理学
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纳米生物学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2024
·Vol. 25
·期 2
·1153
·
IF 4.9
Hongxue Gao,Zhiqiang Liu,Fengrui Song,Junpeng Xing,Zhong Zheng,Zong Hou,Shu Liu
Cell membrane chromatography (CMC) has been widely recognized as a highly efficient technique for in vitro screening of active compounds. Nevertheless, conventional CMC approaches suffer from a restricted repertoire of cell membrane proteins, making them susceptible to oversaturation. Moreover, the binding mechanism between silica gel and proteins primarily relies on intermolecular hydrogen bonding, which is inherently unstable and somewhat hampers the advancement of CMC. Consequently, this investigation aimed to establish a novel CMC column that could augment protein loading, enhance detection throughput, and bolster binding affinity through the introduction of covalent bonding with proteins. This study utilizes polydopamine (PDA)-coated silica gel, which is formed through the self-polymerization of dopamine (DA), as the carrier for the CMC column filler. The objective is to construct the HK-2/SiO2-PDA/CMC model to screen potential therapeutic drugs for gout. To compare the quantity and characteristics of Human Kidney-2 (HK-2) cell membrane proteins immobilized on SiO2-PDA and silica gel, the proteins were immobilized on both surfaces. The results indicate that SiO2-PDA has a notably greater affinity for membrane proteins compared to silica gel, resulting in a significant improvement in detection efficiency. Furthermore, a screening method utilizing HK-2/SiO2-PDA/CMC was utilized to identify seven potential anti-gout compounds derived fromPlantago asiaticaL. (PAL). The effectiveness of these compounds was further validated using an in vitro cell model of uric acid (UA) reabsorption. In conclusion, this study successfully developed and implemented a novel CMC filler, which has practical implications in the field.
研究领域:
药理学
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分析化学
·
肾脏病学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY A
·
2016
·
IF 3.926
Qiqin Wang,Kun Peng,Weijia Chen,Zhen Cao,Peijie Zhu,Yumei Zhao,Yuqiang Wang,Haibo Zhou,Zhengjin Jiang
This study described a simple synthetic methodology for preparing biomembrane mimicking monolithic column. The suggested approach not only simplifies the preparation procedure but also improves the stability of double chain phosphatidylcholine (PC) functionalized monolithic column. The physicochemical properties of the optimized monolithic column were characterized by scanning electron microscopy, energy-dispersive X-ray spectrometry, and nano-LC. Satisfactory column permeability, efficiency, stability and reproducibility were obtained on this double chain PC functionalized monolithic column. It is worth noting that the resulting polymeric monolith exhibits great potential as a useful alternative of commercial immobilized artificial membrane (IAM) columns for in vitro predication of drug-membrane interactions. Furthermore, the comparative study of both double chain and single chain PC functionalized monoliths indicates that the presence or absence of glycerol backbone and the number of acyl chains are not decisive for the predictive ability of IAM monoliths on drug-membrane interactions. This novel PC functionalized monolithic column also exhibited good selectivity for a protein mixture and a set of pharmaceutical compounds.
研究领域:
分析化学
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生物医学工程
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药物科学
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产品关联
DOI
PubMed
高影响因子
Journal of Pharmaceutical Analysis
·
2021
·
IF 4.769
Peijie Zhu,Weijia Chen,Qiqin Wang,Huihui Wu,Meng Ruan,Hongwu Wang,Zhengjin Jiang
In this research, a new phospholipid based monolith was fabricated by in situ co-polymerization of 1-dodecanoyl-2-(11-methacrylamidoundecanoyl)- sn -glycero-3-phosphoethanolamine and ethylene dimethacrylate to mimick bio-membrane environment. Excellent physicochemical properties of this novel monolith that were achieved included column efficiency, stability, and permeability. Moreover, the biomimetic monolith showed outstanding separation capability for a series of intact proteins and small molecules. In particular, it exhibited good potential as an alternative to the commercial immobilized artificial membrane (IAM) column (IAM.PC.DD2) for studying drug-membrane interactions. This study not only enriched the types of IAM stationary phases, but also provided a simple model for the prediction of phosphatidylethanolamine related properties of drug candidates.
研究领域:
分析化学
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药物化学
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生物医学工程
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DOI
PubMed
高影响因子
JOURNAL OF SEPARATION SCIENCE
·
2017
·Vol. 40
·期 22
·4311-4319
·
IF 2.557
Nani Wang,Qiaoyan Zhang,Hailiang Xin,Dan Shou,Luping Qin
A method using osteoblast membrane chromatography coupled with liquid chromatography and time-of-flight mass spectrometry was developed to recognize and identify the specific active components from traditional Chinese medicines. Primary rat osteoblasts were used for the preparation of the stationary phase in the cell chromatography method. Retention components from the cell chromatography were collected and analyzed by liquid chromatography with time-of-flight mass spectrometry. This method was applied in screening active components from extracts of four traditional Chinese medicines. In total, 24 potentially active components with different structures were retained by osteoblast cell chromatography. There were five phenolic glucosides and one triterpenoid saponin from Curculigo orchioides Gaertn, two organic acids and ten flavonoids from Epimedium sagittatum Maxim, one phthalide compound and one organic acid from Angelica sinensis Diels, and two flavonoids and two saponins from Anemarrhena asphodeloides Bunge. Among those, four components (icariin, curculigoside, ferulaic acid, and timosaponin BII) were used for in vitro pharmacodynamics validation. They significantly increased the osteoblast proliferation, alkaline phosphatase activity, levels of bone gla protein and collagen type 1, and promoted mineralized nodule formation. The developed method was an effective screening method for finding active components from complex medicines that act on bone diseases.
研究领域:
药理学
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分析化学
·
中医药
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF SEPARATION SCIENCE
·
2020
·Vol. 43
·期 11
·2105-2114
·
IF 2.878
Can Wu,Nani Wang,Pingcui Xu,Xuping Wang,Dan Shou,Yan Zhu
Cell membrane chromatography is a powerful tool for screening active components from complex matrices. New cell membrane carriers need to be developed to increase the coverage of cell membranes on the surface of stationary phases, thereby improving cell membrane chromatographic retention. In this work, we prepared polyvinyl alcohol-poly dimethyl diallyl ammonium chloride modified silica gel as a cell membrane carrier. Osteoblast cell was used as cell membrane source, which was widely used to evaluate the osteogenic activity of drugs. The new cell membrane chromatographic stationary phase was used to screen anti-osteoporosis components in Liuwei Dihuang decoction-containing serum. The chemical structures of the new modified materials were characterized by scanning electronic microscopy, Fourier transform infrared spectroscopy, and elemental analysis characterization. Compared with the common cell membrane column, the cell membrane coverage of this modified material was increased by 30%, and thus provided a stronger retention effect in positive drugs. Nineteen metabolites in rat serum samples were retained on the cell membrane chromatography and identified by ultra-high-performance liquid chromatography/time-of-flight mass spectrometry. Among those, four components (morroniside, catalpol, loganin, and acteoside) were selected for in vitro pharmacodynamics validation. They significantly increased the osteoblast proliferation. The new modified material was successfully applied to screen anti-osteoporosis components from Liuwei Dihuang Decoction-containing serum.
研究领域:
药学
·
生物技术
·
色谱学
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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）