Analytical biochemistry最新文献_第3页

Production and characterization of a panel of anti-mouse placenta-specific protein 1 (plac1) monoclonal antibodies 抗小鼠胎盘特异性蛋白 1（placenta-specific Protein 1，plac1）单克隆抗体的制备与表征。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-09-26 DOI: 10.1016/j.ab.2024.115682

Sahar Mortezagholi , Faezeh Maghsood , Sorour Shojaeian , Fazel Shokri , Mohammad Mehdi Amiri , Ahmad Ghorbani , Mahdi Shabani , Amir-Hassan Zarnani

{"title":"Production and characterization of a panel of anti-mouse placenta-specific protein 1 (plac1) monoclonal antibodies","authors":"Sahar Mortezagholi , Faezeh Maghsood , Sorour Shojaeian , Fazel Shokri , Mohammad Mehdi Amiri , Ahmad Ghorbani , Mahdi Shabani , Amir-Hassan Zarnani","doi":"10.1016/j.ab.2024.115682","DOIUrl":"10.1016/j.ab.2024.115682","url":null,"abstract":"<div><div>Placenta-Specific Protein 1 (PLAC1) is essential for normal placental and embryonic development. It is widely expressed in various types of cancer cells. We produced a panel of anti-mouse plac1 monoclonal antibodies (mAbs) with different applications. Two recombinant proteins were produced containing either the extracellular domain (ED) plus tetanus toxin P2, P30, pan-DR epitope (PADRE), and KDEL3 (main plac1) or ED plus KDEL3 (control plac1). Recombinant proteins were used for immunization and screening. Positive clones were selected by ELISA and flow cytometry. Purified mAbs were tested by ELISA, WB, flow cytometry, immunohistochemistry (IHC), and immunofluorescent (IF). A combination of bioinformatics tools was used to predict the target epitope(s) of the mAbs. Eight anti-mouse plac1 mAbs (all IgG1/κ1) were generated, all reacting with high affinity in ELISA. Seven clones recognized plac1 in both reduced and non-reduced Western blots, while one only recognized the non-reduced form. Cross-inhibition ELISA revealed that all mAbs recognized overlapping epitopes with a shared motif except for 5C9. Four clones reacted with the native antigen in flow cytometry, but none were functional in IF or IHC staining. The produced multifunctional mAbs can be used to investigate different aspects of PLAC1 biology in reproduction and cancer.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142339518","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}

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Competitive SPR chaser assay to study biomolecular interactions with very slow binding dissociation rate constant 竞争性 SPR 追踪器测定法，用于研究结合解离速率常数极慢的生物分子相互作用。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-09-26 DOI: 10.1016/j.ab.2024.115679

Aye Myat Myat Thinn , Wei Wang , Qing Chen

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Proximity ligation-triggered DNAzyme for selective fluorescent aptasensing of methicillin-resistant Staphylococcus aureus 用于耐甲氧西林金黄色葡萄球菌选择性荧光诱导的近接触发 DNA 酶。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-09-25 DOI: 10.1016/j.ab.2024.115683

Li Wan , Li Feng , Meiling Wang , Yanhui Yang , Pinxiu Pan , Shuhua Gao

{"title":"Proximity ligation-triggered DNAzyme for selective fluorescent aptasensing of methicillin-resistant Staphylococcus aureus","authors":"Li Wan , Li Feng , Meiling Wang , Yanhui Yang , Pinxiu Pan , Shuhua Gao","doi":"10.1016/j.ab.2024.115683","DOIUrl":"10.1016/j.ab.2024.115683","url":null,"abstract":"<div><div>There is an urgent need for novel strategies to accurately and reliably detect pathogenic bacteria to address the global epidemic of antibiotic resistance. This study proposes an innovative approach combining dual aptamer-based target recognition and proximity ligation assay (PLA) triggered DNAzyme recycling cleavage. This method allows for the precise identification and reliable detection of methicillin-resistant <em>Staphylococcus aureus</em> (MRSA). The fluorescence probe labeled with a fluorophore is modified on gold nanoparticles (AuNPs), resulting in the quenching of the fluorescent signal by the AuNPs. The interaction between MRSA and two aptamers leads to forming a Mg<sup>2+</sup>-dependent DNAzyme. The DNAzyme cleaves the fluorescence probe, causing the fluorescent fragment to detach from the surface of the AuNPs, in which the quenched fluorescence signal in the fluorescence probe reappears. The DNAzyme-assisted cleavage and rebinding process generates a processive strolling along the surface track of AuNPs. Consequently, the fluorescence intensity experiences a substantial recovery. A strong linear correlation is observed between the fluorescence intensity and MRSA concentration within 50 cfu/mL to 10<sup>6</sup> cfu/mL. We believe that implementing the novel integrated strategy will broaden the range of bacterial detection methods in the battlefield environment and stimulate the creation of potential new drugs in the future.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142339519","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}

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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

Micaela Riscado , Rita Carapito , Cláudio J. Maia , Chantal Pichon , Mara G. Freire , Mattia Sponchioni , Fani Sousa

{"title":"A new approach for extracellular RNA recovery from Rhodovulum sulfidophilum","authors":"Micaela Riscado , Rita Carapito , Cláudio J. Maia , Chantal Pichon , Mara G. Freire , Mattia Sponchioni , Fani Sousa","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}

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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

Miaowen Sun, Yanzhu Liu, Lingge Shi, Haomiao Dou, Feiyan Ma, Guangda Xu, Longshan Zhao

{"title":"A novel ratiometric colorimetric sensor for detecting hypochlorite and ascorbic acid based on cascade reaction","authors":"Miaowen Sun, Yanzhu Liu, Lingge Shi, Haomiao Dou, Feiyan Ma, Guangda Xu, Longshan Zhao","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}

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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

Nanfei Yang , Yuncheng Bei , Yahong Huang , Wei Zheng , Jiehua Ma , Jiangqiong Ke

{"title":"An electrochemical immunosensor for sensitive and rapid detection of cystatin C based on Fe3O4/AuNPs-MWCNTs@PDA nanocomposite","authors":"Nanfei Yang , Yuncheng Bei , Yahong Huang , Wei Zheng , Jiehua Ma , Jiangqiong Ke","doi":"10.1016/j.ab.2024.115677","DOIUrl":"10.1016/j.ab.2024.115677","url":null,"abstract":"<div><div>Serum Cystatin C (CysC) is an impressive marker for early diagnosis of renal dysfunction. In this work, we established a novel electrochemical immunosensor based on Fe<sub>3</sub>O<sub>4</sub>/AuNPs-MWCNTs@PDA nanocomposite for the detection of CysC. The Fe<sub>3</sub>O<sub>4</sub>/AuNPs-MWCNTs@PDA nanozyme complex by polydopamine encapsulation can not only carry massive detection antibodies, but also bind the electroactive substance toluidine blue (TB) through electrostatic adsorption. By immobilizing AuNPs onto the electrode to bind the capture antibody (Ab1), we constructed a sandwich electrochemical immunosensor with low cost, high sensitivity, and repeatability. The detection range is 3.9–125.0 ng/mL with a significant linear relationship between the current peak difference (ip) and logarithm of the CysC concentration. Moreover, the detection limit of the immunosensor is 0.157 ng/mL. We have successfully utilized this novel immunosensor to detect CysC in human serum samples, and these results have implications for its potential use in clinical application.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142279341","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}

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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

Karthikeyan Murugesan , Marimuthu Dhinesh Kumar , Ganesan Kaniraja , Periyasamy Ananthappan , Vairathevar Sivasamy Vasantha , Chandran Karunakaran

{"title":"Theoretical screening and electrochemical sensor for determination of norepinephrine using a molecularly imprinted poly (3-amiophenylboronic acid)","authors":"Karthikeyan Murugesan , Marimuthu Dhinesh Kumar , Ganesan Kaniraja , Periyasamy Ananthappan , Vairathevar Sivasamy Vasantha , Chandran Karunakaran","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}

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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

Selva Bilge , Manolya Müjgan Gürbüz , Sibel A. Ozkan , Burcu Dogan Topal

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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

Tao Han , Cheng Zhang , Hui Yu , Jinghong Li

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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

Ânderson Ramos Carvalho , Luana Candice Genz Bazana , Marco Flôres Ferrão , Alexandre Meneghello Fuentefria

{"title":"Unraveling the complexities of antifungal susceptibility testing in Candida spp.: Insights from design of experiments","authors":"Ânderson Ramos Carvalho , Luana Candice Genz Bazana , Marco Flôres Ferrão , Alexandre Meneghello Fuentefria","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}

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