Analytical biochemistry最新文献

A new approach for extracellular RNA recovery from Rhodovulum sulfidophilum 从嗜硫红藻中回收细胞外 RNA 的新方法

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-09-24 DOI: 10.1016/j.ab.2024.115681

{"title":"A new approach for extracellular RNA recovery from Rhodovulum sulfidophilum","authors":"","doi":"10.1016/j.ab.2024.115681","DOIUrl":"10.1016/j.ab.2024.115681","url":null,"abstract":"<div><div>The development of RNA-based drugs is highly pursued due to the possibility of creating viable and effective therapies. However, their translation to clinical practice strongly depends on efficient technologies to produce substantial levels of these biomolecules, with high purity and high quality. RNAs are commonly produced by chemical or enzymatic methods, displaying these limitations. In this sense, recombinant production arises as a promising, cost-effective method, allowing large-scale production of RNA. <em>Rhodovulum sulfidophilum (R. sulfidophilum)</em>, a marine purple bacterium, offers the advantage of RNA secretion into the extracellular medium, which contains low levels of RNases and other impurities. Therefore, RNA recovery can be simplified compared to standard extraction protocols involving cell lysis, resulting in a more clarified sample and an improved downstream process. In this work, <em>R. sulfidophilum</em> was transformed with a plasmid DNA encoding pre-miR-29b-1, which is known to be involved in the Alzheimer's disease pathway. After production, the pre-miR-29b-1 was recovered through different extraction methods to verify the most advantageous one. A protocol for extracellular RNA recovery was then established, revealing to be a simpler and less time-consuming method, reducing around 16 h in execution time and the quantity of reagents needed when compared to other established methods. The new strategy brings the additional advantage of eliminating the toxic organic compounds routinely used in intracellular RNA extractions. Overall, the optimized process described here, using isopropanol as the precipitation agent, offers a greener, safer, and faster alternative for recombinant RNA recovery and concentration, with an extracellular RNA recovery of 7 μg/mL and target pre-miRNA-29b-1 recovery of 0.7 μg/L of medium.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142327122","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A novel ratiometric colorimetric sensor for detecting hypochlorite and ascorbic acid based on cascade reaction 基于级联反应检测次氯酸盐和抗坏血酸的新型比率比色传感器

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-09-23 DOI: 10.1016/j.ab.2024.115678

{"title":"A novel ratiometric colorimetric sensor for detecting hypochlorite and ascorbic acid based on cascade reaction","authors":"","doi":"10.1016/j.ab.2024.115678","DOIUrl":"10.1016/j.ab.2024.115678","url":null,"abstract":"<div><div>Hypochlorite and ascorbic acid (AA), play an indispensable role in numerous physiological activities. Herein, a ratiometric colorimetric sensing strategy for the determination of hypochlorite and AA was developed via the catalytic oxidation and reduction of 3,3′,5,5′-tetramethylbenzidine (TMB). Interestingly, in the presence of Fe<sub>3</sub>O<sub>4</sub>-MOF-5(Fe) and hypochlorite, TMB complexes in acidic environments were oxidized to blue oxidized TMB and further diazotized to produce yellow-green diazotized TMB, resulting in the hypochlorite concentration-dependent ratiometric variation for the absorbance at 652 and 450 nm (A<sub>450</sub>/A<sub>652</sub>). Moreover, the diazotized TMB was restored to colorless TMB due to the reducibility of AA, and the detection limit of hypochlorite and AA were 0.027 and 0.677 μM, respectively. The ratiometric colorimetric sensing platform offered higher sensitivity and better selectivity because of the specific hypochlorite-induced reaction and the excellent peroxidase-like activity of Fe<sub>3</sub>O<sub>4</sub>-MOF-5(Fe). The proposed novel strategy provided the guidance to develop sensors for successive detection of hypochlorite and AA in complicated samples.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142320143","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

An electrochemical immunosensor for sensitive and rapid detection of cystatin C based on Fe3O4/AuNPs-MWCNTs@PDA nanocomposite 一种基于 Fe3O4/AuNPs-MWCNTs@PDA 纳米复合材料的电化学免疫传感器，用于灵敏快速地检测胱抑素 C。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-09-21 DOI: 10.1016/j.ab.2024.115677

引用次数: 0

Theoretical screening and electrochemical sensor for determination of norepinephrine using a molecularly imprinted poly (3-amiophenylboronic acid) 使用分子印迹聚（3-氨基苯硼酸）测定去甲肾上腺素的理论筛选和电化学传感器。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-09-20 DOI: 10.1016/j.ab.2024.115676

{"title":"Theoretical screening and electrochemical sensor for determination of norepinephrine using a molecularly imprinted poly (3-amiophenylboronic acid)","authors":"","doi":"10.1016/j.ab.2024.115676","DOIUrl":"10.1016/j.ab.2024.115676","url":null,"abstract":"<div><div>Norepinephrine (NE) is the primary catecholamine (CA) of interest in the medical field, as it plays a key role in regulating the hormonal and neurological systems. Some NE concentration dysfunction can lead to a number of serious physical conditions. As a result, quick and sensitive NE detection is most critical in medical technology. Thus, in this research, a molecularly imprinted polymer (MIP) was used to create an electrochemical sensor for the selective detection of NE. Prior to this, functional monomers were chosen through molecular modeling utilizing molecular mechanics and quantum mechanics computations. According to these studies, the 3-aminophenylboronic acid (3-APBA) functional monomer produces the most stable complex with NE in molecular modeling calculations. Based on this, by electropolymerizing 3-APBA in the presence of the template molecule NE, an imprinting polymer film is formed on the screen-printed carbon electrode (SPCE) surface. Stepwise fabrication of imprinted polymer films was examined through differential pulse voltammetry (DPV), cyclic voltammetry (CV), scanning electron microscopy (SEM), and electrochemical impedance spectroscopy (EIS). The performance of the electrochemical NE sensor removal and rebinding levels of the template was studied and optimized. The selectivity for NE was confirmed by using interference studies of small molecules like dopamine, tyrosine, and serotonin. Under optimum levels, the fabricated MIP sensor had a broad linear range over NE concentrations of 0.1 pM–5 pM; sensitivity: 0.004 mA pM<sup>−1</sup>; limit of detection: 0.03 pM. It is noteworthy that the newly created MIP sensor was effectively validated for NE detection in plasma samples.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142279342","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Electrochemical sensor for the analysis of 5-hydroxymethylcytosine in the presence of cytosine using pencil graphite electrode 使用铅笔石墨电极分析存在胞嘧啶的 5-羟甲基胞嘧啶的电化学传感器

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-09-16 DOI: 10.1016/j.ab.2024.115674

引用次数: 0

A disposable fiber-optic plasmonic sensor for chemical sensing 用于化学传感的一次性光纤质子传感器

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-09-16 DOI: 10.1016/j.ab.2024.115672

引用次数: 0

Unraveling the complexities of antifungal susceptibility testing in Candida spp.: Insights from design of experiments 揭示念珠菌抗真菌药敏试验的复杂性：实验设计的启示

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-09-14 DOI: 10.1016/j.ab.2024.115675

{"title":"Unraveling the complexities of antifungal susceptibility testing in Candida spp.: Insights from design of experiments","authors":"","doi":"10.1016/j.ab.2024.115675","DOIUrl":"10.1016/j.ab.2024.115675","url":null,"abstract":"<div><p>Our study delved into the intricate dynamics of antifungal susceptibility testing for <em>Candida</em> spp., employing a Design of Experiments approach. We systematically investigated the influence of pH, temperature, inoculum size, and glucose concentration on both growth patterns and inhibitory concentrations of <em>Candida</em> spp. Our findings underscore the nuanced interplay between these factors, revealing significant impacts on susceptibility outcomes. Notably, even minor adjustments in these parameters yielded substantial variations in growth and inhibitory concentrations, underscoring the critical importance of meticulous control over growth conditions in antifungal susceptibility testing protocols. Each <em>Candida</em> isolates exhibited unique susceptibility profiles, necessitating tailored culture conditions for accurate testing. Our study sheds light on the variability inherent in <em>Candida</em> spp. growth patterns and emphasizes the need for standardized protocols to ensure consistency across laboratories. By leveraging the design of experiments, our research provides a systematic framework for unraveling the complexities of antifungal susceptibility testing, offering valuable insights for optimizing testing protocols and informing clinical decision-making in antifungal treatment. These findings represent a significant step towards enhancing the efficacy and reliability of antifungal susceptibility testing in clinical practice.</p></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142238574","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Separation of recombinant erythropoietin and human serum albumin without the use of sophisticated equipment 无需复杂设备即可分离重组促红细胞生成素和人血清白蛋白

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-09-13 DOI: 10.1016/j.ab.2024.115673

引用次数: 0

Electrocatalytic efficiency of carbon nitride supported gold nanoparticle based sensor for iodide and cysteine detection 基于氮化碳支撑金纳米粒子的碘化物和半胱氨酸检测传感器的电催化效率

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-09-10 DOI: 10.1016/j.ab.2024.115660

{"title":"Electrocatalytic efficiency of carbon nitride supported gold nanoparticle based sensor for iodide and cysteine detection","authors":"","doi":"10.1016/j.ab.2024.115660","DOIUrl":"10.1016/j.ab.2024.115660","url":null,"abstract":"<div><p>Extensive investigations are being conducted on gold nanoparticles focusing on their applications in biosensors, laser phototherapy, targeted drug delivery and bioimaging utilizing advanced detection techniques. In this work, an electrochemical sensor was developed based on graphite carbon nitride supported gold nanoparticles. Carbon nitride supported gold nanoparticles (Au–CN) was synthesized by applying a deposition-precipitation route followed by a chemical reduction technique. The composite system was characterized by X-ray diffraction and X-ray photo electron spectroscopy methods. Electron microscopy analysis confirmed the formation of gold nanoparticles within the size range of 5–15 nm on the carbon nitride support. Carbon nitride supported gold based sensor was employed for the electrochemical detection of iodide ion and <span>l</span>-cysteine. The limit of detection and sensitivity of the sensor was attained 8.9 μM and 0.96 μAμM⁻<sup>1</sup>cm⁻<sup>2</sup>, respectively, for iodide ion, while 0.48 μM and 5.8 μAμM⁻<sup>1</sup>cm⁻<sup>2</sup>, respectively, was achieved for the recognition of cysteine. Furthermore, a paper-based electrochemical device was developed using the Au–CN hybrid system that exhibited promising results in detecting iodide ions, highlighting its potential for economic and portable device applications.</p></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142168150","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development of a double-antibody sandwich ELISA for quantification of mutated EPSPS gene expression in rice 开发双抗体夹心酶联免疫吸附测定法，用于定量检测水稻中突变的 EPSPS 基因表达量

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-09-10 DOI: 10.1016/j.ab.2024.115669

{"title":"Development of a double-antibody sandwich ELISA for quantification of mutated EPSPS gene expression in rice","authors":"","doi":"10.1016/j.ab.2024.115669","DOIUrl":"10.1016/j.ab.2024.115669","url":null,"abstract":"<div><p>Glyphosate resistance is a critically important trait for genetically modified (GM) crops. Mutation of the rice EPSPS gene results in a high level of glyphosate resistance, presenting significant potential for the development of glyphosate-tolerant crops. The resistance of rice to glyphosate is correlated with the expression levels of resistance genes. Therefore, developing a convenient, stable, and sensitive method for quantifying the OsmEPSPS protein is crucial for the development of glyphosate-resistant crops. We developed a double-antibody sandwich quantitative ELISA (DAS-ELISA) using a specific monoclonal antibody (mAb) for OsmEPSPS capture and an HRP-conjugated <em>anti</em>-OsmEPSPS rabbit polyclonal antibody (pAb). The method could be used to detect OsmEPSPS within a linear range of 16–256 ng/mL with robust intra- and inter-batch duplicability (%CV values: 0.17 %–7.24 %). OsmEPSPS expression in the transgenic rice lines (54.44–445.80 μg/g) was quantified using the DAS-ELISA. Furthermore, the expression of the <em>OsmEPSPS</em> gene was validated through Western blotting. This study demonstrated the reliability and stability of the DAS-ELISA for OsmEPSPS detection in GM rice.</p></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142229085","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0