Analytical biochemistry最新文献_第10页

A fluorescence anisotropy-based competition assay to identify inhibitors against ricin and Shiga toxin ribosome interactions 基于荧光各向异性的竞争测定法，用于识别蓖麻毒素和志贺毒素核糖体相互作用的抑制剂。

IF 2.9 4区生物学

Analytical biochemistry Pub Date : 2024-05-31 DOI: 10.1016/j.ab.2024.115580

Arkajyoti Dutta , Zoltan Szekely , Hakan Guven , Xiao-Ping Li , John E. McLaughlin , Nilgun E. Tumer

{"title":"A fluorescence anisotropy-based competition assay to identify inhibitors against ricin and Shiga toxin ribosome interactions","authors":"Arkajyoti Dutta , Zoltan Szekely , Hakan Guven , Xiao-Ping Li , John E. McLaughlin , Nilgun E. Tumer","doi":"10.1016/j.ab.2024.115580","DOIUrl":"10.1016/j.ab.2024.115580","url":null,"abstract":"<div><p>Ricin is one of the most toxic substances known and a type B biothreat agent. Shiga toxins (Stxs) produced by <em>E. coli</em> (STEC) and <em>Shigella dysenteriae</em> are foodborne pathogens. There is no effective therapy against ricin or STEC and there is an urgent need for inhibitors. Ricin toxin A subunit (RTA) and A1 subunit of Stx2a (Stx2A1) bind to the C-terminal domain (CTD) of the ribosomal P-stalk proteins to depurinate the sarcin/ricin loop. Modulation of toxin-ribosome interactions has not been explored as a strategy for inhibition. Therefore, development of assays that detect inhibitors targeting toxin-ribosome interactions remains a critical need. Here we describe a fluorescence anisotropy (FA)-based competitive binding assay using a BODIPY-TMR labeled 11-mer peptide (P11) derived from the P-stalk CTD to measure the binding affinity of peptides ranging from 3 to 11 amino acids for the P-stalk pocket of RTA and Stx2A1. Comparison of the affinity with the surface plasmon resonance (SPR) assay indicated that although the rank order was the same by both methods, the FA assay could differentiate better between peptides that show nonspecific interactions by SPR. The FA assay detects only interactions that compete with the labeled P11 and can validate inhibitor specificity and mechanism of action.</p></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0003269724001246/pdfft?md5=a6c263d17695c84cc0f06483b236bcd5&pid=1-s2.0-S0003269724001246-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141199154","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

High-level expression of codon-optimized Taq DNA polymerase under the control of rhaBAD promoter 在 rhaBAD 启动子控制下高水平表达代码优化的 Taq DNA 聚合酶。

IF 2.9 4区生物学

Analytical biochemistry Pub Date : 2024-05-28 DOI: 10.1016/j.ab.2024.115581

Fina Amreta Laksmi , Kartika Sari Dewi , Isa Nuryana , Siti Eka Yulianti , Kharisma Panji Ramadhan , Moch Irfan Hadi , Yudhi Nugraha

{"title":"High-level expression of codon-optimized Taq DNA polymerase under the control of rhaBAD promoter","authors":"Fina Amreta Laksmi , Kartika Sari Dewi , Isa Nuryana , Siti Eka Yulianti , Kharisma Panji Ramadhan , Moch Irfan Hadi , Yudhi Nugraha","doi":"10.1016/j.ab.2024.115581","DOIUrl":"10.1016/j.ab.2024.115581","url":null,"abstract":"<div><p>A DNA polymerase from <em>Thermus aquaticus</em> remains the most popular among DNA polymerases. It was widely applied in various fields involving the application of polymerase chain reaction (PCR), implying the high commercial value of this enzyme. For this reason, an attempt to obtain a high yield of Taq DNA polymerase is continuously conducted. In this study, the <span>l</span>-rhamnose-inducible promoter rhaBAD was utilized due to its ability to produce recombinant protein under tight control in <em>E. coli</em> expression system. Instead of full-length Taq polymerase, an N-terminal deletion of Taq polymerase was selected. To obtain a high-level expression, we attempted to optimize the codon by reducing the rare codon and GC content, and in a second attempt, we optimized the culture conditions for protein expression. The production of Taq polymerase using the optimum culture condition improved the level of expression by up to 3-fold. This approach further proved that a high level of recombinant protein expression could be achieved by yielding a purified Taq polymerase of about 8.5 mg/L of culture. This is the first research publication on the production of Taq polymerase with N-terminal deletion in <em>E. coli</em> with the control of the rhaBAD promoter system.</p></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141178833","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

Recent advancements in non-invasive wearable electrochemical biosensors for biomarker analysis – A review 用于生物标记分析的非侵入式可穿戴电化学生物传感器的最新进展 - 综述。

IF 2.9 4区生物学

Analytical biochemistry Pub Date : 2024-05-25 DOI: 10.1016/j.ab.2024.115578

G. Backiyalakshmi , U. Snekhalatha , Anela L. Salvador

{"title":"Recent advancements in non-invasive wearable electrochemical biosensors for biomarker analysis – A review","authors":"G. Backiyalakshmi , U. Snekhalatha , Anela L. Salvador","doi":"10.1016/j.ab.2024.115578","DOIUrl":"10.1016/j.ab.2024.115578","url":null,"abstract":"<div><p>A biomarker is a molecular indicator that can be used to identify the presence or severity of a disease. It may be produced due to biochemical or molecular changes in normal biological processes. In some cases, the presence of a biomarker itself is an indication of the disease, while in other cases, the elevated or depleted level of a particular protein or chemical substance aids in identifying a disease. Biomarkers indicate the progression of the disease in response to therapeutic interventions. Identifying these biomarkers can assist in diagnosing the disease early and providing proper therapeutic treatment. In recent years, wearable electrochemical (EC) biosensors have emerged as an important tool for early detection due to their excellent selectivity, low cost, ease of fabrication, and improved sensitivity. There are several challenges in developing a fully integrated wearable sensor, such as device miniaturization, high power consumption, incorporation of a power source, and maintaining the integrity and durability of the biomarker for long-term continuous monitoring. This review covers the recent advancements in the fabrication techniques involved in device development, the types of sensing platforms utilized, different materials used, challenges, and future developments in the field of wearable biosensors.</p></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-05-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141157826","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

Comprehensive overview of analytical and bioanalytical methodologies for the opioid analgesics - Tramadol and combinations 阿片类镇痛药 - 曲马多及其复方制剂的分析和生物分析方法综述

IF 2.9 4区生物学

Analytical biochemistry Pub Date : 2024-05-24 DOI: 10.1016/j.ab.2024.115579

K.S. Kokilambigai , V.M. Irina , K.C. Sheba Mariam , K. Adila , S. Kathirvel

引用次数: 0

Touchpad-based immunochromatographic strip for detecting the skin surface proteins 基于触摸板的免疫层析条带，用于检测皮肤表面蛋白质

IF 2.9 4区生物学

Analytical biochemistry Pub Date : 2024-05-23 DOI: 10.1016/j.ab.2024.115575

Hyugo Tsukada , Mai Wako , Syunsuke Ueda , Kuniaki Nagamine

{"title":"Touchpad-based immunochromatographic strip for detecting the skin surface proteins","authors":"Hyugo Tsukada , Mai Wako , Syunsuke Ueda , Kuniaki Nagamine","doi":"10.1016/j.ab.2024.115575","DOIUrl":"10.1016/j.ab.2024.115575","url":null,"abstract":"<div><p>This study demonstrates, for the first time, the proof-of-concept of a novel immunosensor, a touchpad-based immunochromatographic strip, that non-invasively extracts and detects skin surface proteins. The strip was composed of a nitrocellulose membrane at the center, where a spot of anti-human IgG capture antibody was physically adsorbed. The capture antibody spot was covered with a glass fiber membrane impregnated with phosphate-buffered saline (PBS) to extract skin surface proteins, avoiding direct contact of the human skin with the capture antibodies. Skin surface IgG was detected in two steps: (1) touching the capture antibody via a glass fiber membrane containing PBS, and (2) dipping the strip into the Au-nanoparticle-labeled secondary antibody to visualize the existence of the captured skin surface IgG on the strip. We qualitatively demonstrated that using a very small amount of PBS while maintaining contact with the skin, skin surface proteins can be concentrated and detected, even with a relatively low-sensitivity immunochromatographic chip. This sensor is expected to be a potential biosensor for the non-invasive diagnosis of the integrity of human skin.</p></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141133266","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

Validation of one-step reverse transcription digital PCR assays for Norovirus GI 一步式反转录数字 PCR 检测诺如病毒 GI 的验证

IF 2.9 4区生物学

Analytical biochemistry Pub Date : 2024-05-23 DOI: 10.1016/j.ab.2024.115576

Bomin Ko , Taejin Shin , Boram Kim , Da-Hye Lee

引用次数: 0

Electrochemical detection of myoglobin using an ultrasensitive label-free sensor derived from cubic-ZIF67@Au-rGOF-NH2 composite of MOF and GOF 利用 MOF 和 GOF 的立方体-ZIF67@Au-rGOF-NH2 复合材料衍生的超灵敏无标记传感器电化学检测肌红蛋白

IF 2.9 4区生物学

Analytical biochemistry Pub Date : 2024-05-23 DOI: 10.1016/j.ab.2024.115571

Chunyan Wu , Wendi Mu , Fangfang Wu , Hongmin Gao , Xinshui Ren , Jing Feng , Men Miao , Hehua Zhang , Dong Chang , Hongzhi Pan

{"title":"Electrochemical detection of myoglobin using an ultrasensitive label-free sensor derived from cubic-ZIF67@Au-rGOF-NH2 composite of MOF and GOF","authors":"Chunyan Wu , Wendi Mu , Fangfang Wu , Hongmin Gao , Xinshui Ren , Jing Feng , Men Miao , Hehua Zhang , Dong Chang , Hongzhi Pan","doi":"10.1016/j.ab.2024.115571","DOIUrl":"10.1016/j.ab.2024.115571","url":null,"abstract":"<div><p>Markers of myocardial injury, such as myoglobin (Mb), are substances swiftly released into the peripheral bloodstream upon myocardial cell injury or altered cardiac activity. During the onset of acute myocardial infarction, patients experience a significant surge in serum Mb levels. Given this, precise detection of Mb is essential, necessitating the development of innovative assays to optimize detection capabilities. This study introduces the synthesis of a three-dimensional hierarchical nanocomposite, Cubic-ZIF67@Au-rGOF-NH<sub>2</sub>, utilizing aminated reduced graphene oxide and zeolite imidazolium ester framework-67 (ZIF67) as foundational structures. Notably, this novel material, applied in a label-free electrochemical immunosensor, presents a groundbreaking approach for detecting myocardial injury markers. Experimental outcomes revealed ZIF67 and AuNPs exhibit enhanced affinity and growth on the 3D-rGOF-NH<sub>2</sub> matrix, thus amplifying electrical conductivity while preserving the inherent electrochemical attributes of ZIF67. As a result, the Cubic-ZIF67@Au-rGOF-NH<sub>2</sub> label-free electrochemical immunosensor exhibited a broad detection range and high sensitivity for Mb. The derived standard curve was ΔIp = 16.67552lgC+275.245 (R = 0.993) with a detection threshold of 3.47 fg/ml. Moreover, recoveries of standards spiked into samples ranged between 96.3% and 108.7%. Importantly, the devised immunosensor retained notable selectivity against non-target proteins, proving its potential clinical utility based on exemplary sample analysis performance.</p></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141140937","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

CeO2·ZnO@biomass-derived carbon nanocomposite-based electrochemical sensor for efficient detection of ascorbic acid 基于 CeO2.ZnO@ 生物质衍生碳纳米复合材料的电化学传感器用于高效检测抗坏血酸。

IF 2.9 4区生物学

Analytical biochemistry Pub Date : 2024-05-22 DOI: 10.1016/j.ab.2024.115574

Jahir Ahmed , M. Faisal , Jari S. Algethami , Mabkhoot Alsaiari , Mohammed Jalalah , Farid A. Harraz

{"title":"CeO2·ZnO@biomass-derived carbon nanocomposite-based electrochemical sensor for efficient detection of ascorbic acid","authors":"Jahir Ahmed , M. Faisal , Jari S. Algethami , Mabkhoot Alsaiari , Mohammed Jalalah , Farid A. Harraz","doi":"10.1016/j.ab.2024.115574","DOIUrl":"10.1016/j.ab.2024.115574","url":null,"abstract":"<div><p>Ascorbic acid (AA), a prominent antioxidant commonly found in human blood serum, serves as a biomarker for assessing oxidative stress levels. Therefore, precise detection of AA is crucial for swiftly diagnosing conditions arising from abnormal AA levels. Consequently, the primary aim of this research is to develop a sensitive and selective electrochemical sensor for accurate AA determination. To accomplish this aim, we used a novel nanocomposite comprised of CeO<sub>2</sub>-doped ZnO adorned on biomass-derived carbon (CeO<sub>2</sub>·ZnO@BC) as the active nanomaterial, effectively fabricating a glassy carbon electrode (GCE). Various analytical techniques were employed to scrutinize the structure and morphology features of the CeO<sub>2</sub>·ZnO@BC nanocomposite, ensuring its suitability as the sensing nanomaterial. This innovative sensor is capable of quantifying a wide range of AA concentrations, spanning from 0.5 to 1925 μM in a neutral phosphate buffer solution. It exhibits a remarkable sensitivity of 0.2267 μA μM<sup>−1</sup>cm<sup>−2</sup> and a practical detection limit of 0.022 μM. Thanks to its exceptional sensitivity and selectivity, this sensor enables highly accurate determination of AA concentrations in real samples. Moreover, its superior reproducibility, repeatability, and stability underscore its reliability and robustness for AA quantification.</p></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141086334","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

PicoGreen assay for nucleic acid quantification - Applications, challenges, and solutions 用于核酸定量的 PicoGreen 分析法--应用、挑战和解决方案。

IF 2.9 4区生物学

Analytical biochemistry Pub Date : 2024-05-22 DOI: 10.1016/j.ab.2024.115577

Eunmi Ban, Aeri Kim

{"title":"PicoGreen assay for nucleic acid quantification - Applications, challenges, and solutions","authors":"Eunmi Ban, Aeri Kim","doi":"10.1016/j.ab.2024.115577","DOIUrl":"10.1016/j.ab.2024.115577","url":null,"abstract":"<div><p>Various analytical methods and reagents have been employed for nucleic acid analysis in cells, biological fluids, and formulations. Standard techniques like gel electrophoresis and qRT-PCR are widely used for qualitative and quantitative nucleic acid analysis. However, these methods can be time-consuming and labor-intensive, with limitations such as inapplicability to small RNA at low concentrations and high costs associated with qRT-PCR reagents and instruments. As an alternative, PicoGreen (PG) has emerged as a valuable method for the quantitative analysis of nucleic acids. PG, a fluorescent dye, enables the quantitation of double-stranded DNA (dsDNA) or double-stranded RNA, including miRNA mimic and siRNA, in solution. It is also applicable to DNA and RNA analysis within cells using techniques like FACS and fluorescence microscopy. Despite its advantages, PG's fluorescence intensity is affected by various experimental conditions, such as pH, salts, and chemical reagents. This review explores the recent applications of PG as a rapid, cost-effective, robust, and accurate assay tool for nucleic acid quantification. We also address the limitations of PG and discuss approaches to overcome these challenges, recognizing the expanding range of its applications.</p></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141092726","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

Smartphone-enabled colorimetric immunoassay for deoxynivalenol based on Mn2+-mediated aggregation of AuNPs 基于 Mn2+ 介导的 AuNPs 聚合的智能手机比色法免疫测定脱氧雪腐镰刀菌烯醇。

IF 2.9 4区生物学

Analytical biochemistry Pub Date : 2024-05-20 DOI: 10.1016/j.ab.2024.115572

Xinrui Feng , Qinwei Xu , Yan Liu , Sijia Wang , Yong Cao , Chen Zhao , Shuai Peng

{"title":"Smartphone-enabled colorimetric immunoassay for deoxynivalenol based on Mn2+-mediated aggregation of AuNPs","authors":"Xinrui Feng , Qinwei Xu , Yan Liu , Sijia Wang , Yong Cao , Chen Zhao , Shuai Peng","doi":"10.1016/j.ab.2024.115572","DOIUrl":"10.1016/j.ab.2024.115572","url":null,"abstract":"<div><p>Deoxynivalenol (DON) is a common mycotoxin in food that mainly pollutes grain crops and feeds, such as barley, wheat and corn. DON has caused widespread concern in the field of food and feed safety. In this study, a colorimetric immunoassay was proposed based on the aggregation of gold nanoparticles (AuNPs) due to the decomposition of Mn<sup>2+</sup> from gold-coated manganese dioxide (AuNP@MnO<sub>2</sub>) nanosheets. In this study, 2-(dihydrogen phosphate)-<span>l</span>-ascorbic acid (AAP) was hydrolyzed by alkaline phosphatase (ALP) and converted to ascorbic acid (AA). Then, AuNP@MnO<sub>2</sub> was reduced to Mn<sup>2+</sup> and AuNPs aggregation occurred. Using the unique optical characteristics of AuNPs and AuNP@MnO<sub>2</sub>, visible color changes realized simple detection of DON with high sensitivity and portability. With increasing DON content, the color changed more obviously. To quantitatively detect DON, pictures can be taken and the blue value can be read by a smartphone. The detection limit (Ic10) of this method was 0.098 ng mL<sup>−1</sup>, which was 326 times higher than that of traditional competitive ELISA, and the detection range was 0.177–6.073 ng mL<sup>−1</sup>. This method exhibited high specificity with no cross-reaction in other structural analogs. The average recovery rate of DON in corn flour samples was 89.1 %–110.2 %, demonstrating the high accuracy and stability of this assay in actual sample detection. Therefore, the colorimetric immunoassay can be used for DON-related food safety monitoring.</p></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141080459","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