molecular imprinting technique

  • 文章类型: Journal Article
    纳米酶由于其良好的稳定性和低成本而被认为是生物测定中天然酶的理想替代品。然而,它们在传感中的应用通常具有较差的选择性。例如,Au基纳米酶,作为一种经典的葡萄糖氧化酶模拟酶,可以催化不同的单糖。因此,增强Au基纳米酶的选择性对于构建特定的葡萄糖传感平台具有极大的必要性和紧迫性。在我们的研究中,成功制备了易于回收的聚多巴胺(PDA)支持的Au基纳米酶(PDA@AuNPs),可以催化包括葡萄糖在内的多种单糖,木糖,甘露糖,和蔗糖。为了提高PDA@AuNPs的选择性,以葡萄糖和硼酸衍生物为模板和功能单体,在PDA@AuNPs表面构建了分子印迹聚合物(MIPs)。令人印象深刻的是,获得的分子印迹纳米酶(PDA@AuNPs-MIPs)的催化活性仅显示出轻微的下降(6.3%),而其选择性则明显增强(≥230%)。因此,制备的传感器在10μM-1mM的浓度范围内实现了对葡萄糖的灵敏和选择性检测,低检测限(LOD)为0.227μM(S/N=3),在很大程度上避免了传感溶液中其他单糖的影响。不出所料,准备好的传感器也显示出良好的恢复,和长期稳定。
    Nanozymes have been regarded as the ideal alternatives to natural enzymes in bioassays due to their good stability and low cost. However, their applications in sensing usually suffer from poor selectivity. For example, Au-based nanozymes, as a kind of classical glucose oxidase mimic enzyme, could catalyze diverse monosaccharides. Therefore, it is of great necessity and urgency to endow the Au-based nanozymes with enhanced selectivity for the construction of specific glucose sensing platform. In our study, easily recyclable polydopamine (PDA)-supported Au-based nanozymes (PDA@Au NPs) were successfully prepared and could catalyze diverse monosaccharides including glucose, xylose, mannose, and sucrose. To enhance the selectivity of PDA@Au NPs, molecularly imprinted polymers (MIPs) were constructed on the surface of PDA@Au NPs using glucose and boronic acid derivatives as template and functional monomer. Impressively, the catalytic activity of the obtained molecularly imprinted nanozyme (PDA@Au NPs-MIPs) only shows a slight decrease (6.3%) while their selectivity is obviously enhanced (≥230%). Accordingly, the as-prepared sensor achieved the sensitive and selective detection of glucose in the concentration range of 10 μM-1 mM and a low detection limit (LOD) of 0.227 μM (S/N = 3), avoiding the influence of other monosaccharides exited in the sensing solutions to a great extent. As expected, the as-prepared sensors also showed good recovery, and long-term stability.
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  • 文章类型: Journal Article
    Estrone(E1),作为内源性雌激素,人体具有多种生理功能,对人体健康具有重要意义。另一方面,它是一种广泛分布和高度令人不安的环境内分泌干扰物。因此,迫切需要发展一种敏感的,快速,和便宜的方法,用于现场确定E1,这不仅是临床诊断和治疗,还用于环境水中内源性雌激素污染的调查和监测。在这项研究中,通过表面静电吸附和离子交换制备了Ru(bpy)32+/MWCNTs/Nafion/金电极。通过溶胶-凝胶法制备了具有识别E1分子能力的分子印迹膜(MIP),并用MIP修饰电极以形成电化学发光传感器(MIP-ECL)。该方法同时具有ECL的高灵敏度优势和MIP的高选择性优势。此外,羧化多壁碳纳米管(MWCNT-COOH)的加入改善了金电极表面的功能化并增加了MIP的结合位点。同时,MWCNTs良好的导电性促进了电子转移,进一步提高了传感器的灵敏度。传感器显示了一个宽的线性区间,其中E1浓度范围从0.1μg/L到200μg/L,具有较高的线性相关系数(R2=0.999)。传感器的线性回归方程为Y=243.64x-79.989,检出限(LOD)为0.0047μg/L。为了验证我们的传感器,实际样品也通过参考方法(LC-MS/MS)测量,两种方法定量结果的相对偏差均小于4.1%。这表明该传感器获得的定量结果准确,可用于临床样品和环境水中E1的快速原位测定。
    Estrone (E1), as an endogenous estrogen, has a variety of physiological functions in human body and is of great significance to human health. On the other hand, it is a widely distributed and highly disturbing environmental endocrine disruptor in water. Therefore, there is an urgent need to develop a sensitive, rapid, and inexpensive method for the on-site determination of E1, which is not only for clinical diagnosis and treatment, but also for the investigation and monitoring of endogenous estrogen pollution in environmental water. In this study, Ru(bpy)3 2+/MWCNTs/Nafion/gold electrodes were prepared by surface electrostatic adsorption and ion exchange. A molecularly imprinted membrane (MIP) with the capability to recognize E1 molecules was prepared by sol-gel method, and the electrodes were modified with MIP to form an electrochemical luminescence sensor (MIP-ECL). This method simultaneously possesses ECL\'s advantage of high sensitivity and MIP\'s advantage of high selectivity. Moreover, the addition of carboxylated multi-walled carbon nanotubes (MWCNT-COOH) improved the functionalization of the gold electrode surface and increased the binding sites of MIP. Meanwhile, the good conductivity of MWCNTs promoted electron transfer and further improved the sensitivity of the sensor. The sensor showed a wide linear interval in which the E1 concentrations can range from 0.1 μg/L to 200 μg/L, along with a high linear correlation coefficient (R 2 = 0.999). The linear regression equation of the sensor was Y = 243.64x-79.989, and the detection limit (LOD) was 0.0047 μg/L. To validate our sensor, actual samples were also measured by the reference method (LC-MS/MS), and it was found that the relative deviation of quantitative results of the two different methods was less than 4.1%. This indicates that the quantitative results obtained by this sensor are accurate and can be used for rapid in situ determination of E1 in clinical samples and environmental water.
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  • 文章类型: Journal Article
    这项工作的目的是研究分子印迹技术应用的有效性,以创建用于选择性吸附轻和重稀土金属的高选择性大分子吸附剂(例如,钐和钆,分别),随后彼此分离。这些吸附剂似乎是有希望的,因为在分子印迹聚合物的合成过程中形成了互补的空穴,因此只有目标稀土金属会被吸附。换句话说,所提出的大分子的优点是没有伴随的金属吸附具有密切的化学性质。基于甲基丙烯酸(MAA)和4-乙烯基吡啶(4VP)功能单体合成了两种类型的分子印迹聚合物(MIP)。吸附性能(萃取度,交换能力)的研究。研究了模板去除循环计数(从20到35)对吸附效率的影响。进行了从模型溶液中选择性吸附和分离钐和钆的实验室实验。
    The aim of the work is to study the effectiveness of a molecular imprinting technique application for the creation of highly selective macromolecular sorbents for selective sorption of light and heavy rare-earth metals (for example, samarium and gadolinium, respectively) with subsequent separation from each other. These sorbents seem to be promising due to the fact that only the target rare-earth metal will be sorbed owing to the fact that complementary cavities are formed during the synthesis of molecularly imprinted polymers. In other words, the advantage of the proposed macromolecules is the absence of accompanying sorption of metals with close chemical properties. Two types of molecularly imprinted polymers (MIP) were synthetized based on methacrylic acid (MAA) and 4-vinylpyridine (4VP) functional monomers. The sorption properties (extraction degree, exchange capacity) of the MIPs were studied. The impact of template removal cycle count (from 20 to 35) on the sorption effectivity was studied. Laboratory experiments on selective sorption and separation of samarium and gadolinium from a model solution were carried out.
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  • 文章类型: Journal Article
    Nanozymes have advantages over natural enzymes in terms of efficiency, stability, and economy. MVSM (Mixed Valence State MOF) is a nano-oxidase with uricase-like activity that may catalyze uric acid (UA) in the body into allantoin and H2 O2 to treat gout and hyperuricemia by substituting natural uricase. However, it cannot specifically identify and choose UA. To increase the selectivity and affinity of MVSM for UA, the composite material MVSM@MIP is innovatively synthesized using a new synthetic approach termed the \"two-step synthesis method,\" which may prevent UA from being oxidized by MVSM during manufacture in this study. At the same time, this study also provides experimental proof of the effective creation of the material, the advantages of the \"two-step synthesis approach,\" and the high selectivity and affinity of MVSM@MIP for UA. Based on these findings, the suggested technique may be used to effectively catalyze uric acid in human urine with high activity.
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  • 文章类型: Journal Article
    蛋白A是IgG纯化中最常用的配体,因为它与大多数免疫球蛋白的Fc受体特异性结合,使其在商业上变得重要。分子印迹是一种基于各种分子的选择性识别的方法。分子印迹聚合物是易于制备的材料,耐用,价格便宜,具有分子识别能力。在部分冷冻的环境中通过自由基聚合制备冷冻凝胶。冷凝胶的独特结构与渗透相结合,化学和机械稳定性使它们成为各种生物化合物/标本的有吸引力的色谱基质(质粒,病原体,cells).在这个协议中,蛋白质A印迹的超大孔聚(甲基丙烯酸2-羟乙酯)冷冻凝胶以球形形式制备用于蛋白质A纯化。通过溶胀试验对制备的冷冻凝胶进行表征,扫描电子显微镜(SEM),傅里叶变换红外分光光度计(FTIR),和Brunauer-Emmett-Teller(BET)表面积分析。在表征之后,在分批系统中确定了蛋白质A吸附的最佳条件。在优化印迹冷冻凝胶后确定最大蛋白A吸附能力。针对Fc和蛋白质G检查印迹冷冻凝胶的蛋白质A相对选择性系数。使用快速高效液相色谱(FPLC)从细菌细胞壁分离蛋白质A。通过十二烷基硫酸钠凝胶电泳(SDS-PAGE)测定分离的蛋白A。在最后阶段,检查了冷凝胶的可重用性。
    Protein A is the most commonly used ligand in IgG purification due to its specific binding to the Fc receptor of most immunoglobulins, making it commercially important. Molecular imprinting is a method based on the selective recognition of various molecules. Molecular imprinted polymers are materials that are easy to prepare, durable, cheap and have molecular recognition capability. Cryogels are prepared by radical polymerization in a partially frozen environment. The unique structure of cryogels combined with osmotic, chemical and mechanical stability make them attractive chromatography matrices for a variety of biological compounds/specimens (plasmids, pathogens, cells). In this protocol, protein A imprinted supermacroporous poly(2-hydroxyethyl methacrylate) cryogels were prepared in spherical form for protein A purification. The characterization of the prepared cryogels were made by swelling test, scanning electron microscopy (SEM), Fourier transform infrared spectrophotometer (FTIR), and Brunauer-Emmett-Teller (BET) surface area analysis. After characterization, optimum conditions for protein A adsorption were determined in the batch system. The maximum protein A adsorption capacity was determined after optimization of the imprinted cryogels. Protein A relative selectivity coefficients of imprinted cryogels were examined for both Fc and protein G. Protein A was isolated from the bacterial cell wall using fast performance liquid chromatography (FPLC). The separated protein A was determined by sodium dodecyl sulfate gel electrophoresis (SDS-PAGE). In the last stage, the reusability of the cryogel was examined.
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  • 文章类型: Journal Article
    为了克服低丰度赖氨酸乙酰化(Kac)的鉴定挑战,开发了一种基于分子印迹聚合物(MIP)的新方法,以提高实际样品中Kac肽的提取能力。引入绿色低共熔溶剂(DESs),并将其用作与丙烯酸锌(ZnA)的协同功能单体之一。选择甘氨酸-甘氨酸-丙氨酸-赖氨酸(ac)-精氨酸(GAKacR)作为模板,N'-亚甲基双丙烯酰胺(MBAA)用作交联剂。获得的GGAKacR-MIP对模板具有优异的选择性,印迹因子(IF)高达21.4。组蛋白消化添加实验表明GGAKacR-MIP可以成功地从复杂样品中提取GGAKacR。最后,详细研究了从小鼠肝脏蛋白消化中提取Kac肽的应用。所鉴定的Kac肽和Kac蛋白的数量为130和110,比未处理的样品高3.71倍和3.93倍。此外,处理后鉴定的肽和蛋白质的数量从5535和1092增加到17149和4037(3.10倍和3.70倍,分别)。结果表明,获得的MIP可以为Kac修饰和肽分馏的鉴定提供有效的技术工具,以及同时识别翻译后修饰的蛋白质组和蛋白质组信息的潜在方法。
    To overcome the identification challenge of low-abundance lysine acetylation (Kac), a novel approach based on a molecularly imprinted polymer (MIP) was developed to improve the extraction capacity of Kac peptides in real samples. Green deep eutectic solvents (DESs) were introduced and used as one of the synergistic functional monomers with zinc acrylate (ZnA). Glycine-glycine-alanine-lysine(ac)-arginine (GGAKacR) was chosen as a template and N,N\'-methylenbisacrylamide (MBAA) was used as a cross-linker. The obtained GGAKacR-MIP had excellent selectivity for the template with an imprinting factor (IF) of up to 21.4. The histone digest addition experiment demonstrated that GGAKacR-MIP could successfully extract GGAKacR from a complex sample. Finally, the application to the extraction of Kac peptides from mouse liver protein digestion was studied in detail. The number of Kac peptides and Kac proteins identified was 130 and 110, which were 3.71-fold and 3.93-fold higher than those of the untreated sample. In addition, the number of peptides and proteins identified after treatment increased from 5535 and 1092 to 17 149 and 4037 (3.10-fold and 3.70-fold, respectively). The results showed that the obtained MIP may provide an effective technical tool for the identification of Kac-modification and peptide fractionation, as well as a potential approach for simultaneously identifying post-translational-modified proteomic and proteomic information.
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  • 文章类型: Journal Article
    In this study, poly(3, 4-ethylenedioxythiophene) (PEDOT) nanocluster structure was synthesized on the reduced graphene oxide (rGO) modified cotton fibers. The organic electrochemical transistors based on the modified fiber have been assembled and their performance of different gate electrode transistors has been investigated. The transistor exhibits an excellent transconductance of up to 15.5 mS and a high on-off ratio close to 2*102. The bending angle and bending times have little effect on the device performance. The uric acid (UA) sensor based transistor has been fabricated for the first time. Flexible sensors based on molecularly imprinted polymer (MIP) membrane with different fiber gate electrodes have been investigated. The UA sensor with MIP/PEDOT/carbon fiber as the gate electrode has a sensitivity of 100 μA per decade from 1 nM to 500 μM, a linear coefficient of 0.97143, excellent selectivity, and good reproducibility. In addition, fiber based organic electrochemical transistors (FECTs) can be sewn into the fabric for monitoring and have successfully evaluated the detection of UA in artificial urine sample, with data consistent well with the UA concentration obtained from single fiber. Therefore, the sensor based FECTs can be used for low cost, accurate, non-enzymatic detection of UA in clinical diagnostics and bioanalytical applications.
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  • 文章类型: Journal Article
    Neopterin (Neo) is thought of as a key biomarker for the diagnosis and prognosis of a wide variety of diseases associated with cellular immune response. Therefore, it has become a vital need to be able to specifically determine the Neo concentration in human serum. Molecularly imprinted cryogels have come into prominence among other affinity systems by combining advantages of Molecular Imprinting Technology (MIT) and cryogels. In this chapter, synthesis of novel Neopterin-imprinted cryogel membranes (Neo-mip), characterization studies of synthesized materials, and their use in the determination of Neo in human serum is described in detail. In addition, the evaluation of selective Neo adsorption properties of Neo-mip against competitors (Pterin and Glucose) is discussed. Neo-mip will come into prominence as important affinity materials for the selective Neo recognition in body fluids, prior to use in the health sector.
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  • 文章类型: Journal Article
    A sensitive, rapid, and cost-effective method for quantitative analysis of proteins (e.g., detection, purification, depletion) for a wide variety of purposes is required in a number of areas, such as immunodiagnostics and biotechnology. Double-layer imprinting technique, which is carried out via polymerization of polymer solution with higher monomer concentration, covering and filling the supermacroporous structure of a pre-synthesized cryogel column with a lower monomer concentration, thus improving the surface area and adsorption capacity of final product, is a brand new approach for the application of cryogels in molecular imprinting technology. Within the scope of this chapter, BSA is selected as a model protein for the application of double-layer imprinting protocol. In this chapter, synthesis of double-layer BSA-imprinted and non-imprinted cryogel columns (BSA-DLIP and DLNIP, respectively) are described. In addition, characterization of synthesized columns and BSA depletion studies from aqueous solutions are described in detail, as well as selectivity of BSA-DLIPs for BSA, against competitors.
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  • 文章类型: Journal Article
    In this paper, a novel molecularly imprinted polymer (MIP) for specific adsorption of steviol glycosides was designed, and the imprinting mechanism of self-assembly system between template and monomers was clearly explored. Firstly, steviol (STE) was chosen as dummy template, and the density functional theory (DFT) at B3LYP/6-31 + G (d, p) level was used to select monomers, imprinting molar ratios, solvents, and cross-linking agents. The selectivity to five steviol glycosides was also calculated. Importantly, reduced density gradient (RDG) theory combined with atom in molecules (AIM) and infrared spectrum (IR) was applied to investigate the bonding situation and the nature of noncovalent interaction in self-assembly system. The theoretical designed results showed that the template which interacts with acrylic acid (AA) has the minimum binding energy, and the complex with the molar ratio of 1 : 4 has the most stable structure. Toluene (TL) and ethylene glycol dimethacrylate (EGDMA) were chosen as the optimal solvent and cross-linking agent, respectively. Five hydrogen bonds formed in the self-assembly system are the key forces at the adsorption sites of MIPs through the RDG and AIM analyses. The MIPs were synthesized by theoretical predictions, and the results showed that the maximum adsorption capacity towards dulcoside A is 26.17 mg/g. This work provided a theoretical direction and experimental validation for deeper researches of the MIPs for steviol glycosides. In addition, the method of RDG theory coupled with AIM and IR also could be used to analyze other imprinting formation mechanisms systematically.
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