Analytical biochemistry最新文献_第10页

Detection of Salmonella by competitive ELISA of lipopolysaccharide secreted into the culture medium 通过对分泌到培养基中的脂多糖进行竞争性 ELISA 检测沙门氏菌。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-10-23 DOI: 10.1016/j.ab.2024.115695

Elena Kiseleva, Konstantin Mikhailopulo, Oleg Sviridov

{"title":"Detection of Salmonella by competitive ELISA of lipopolysaccharide secreted into the culture medium","authors":"Elena Kiseleva, Konstantin Mikhailopulo, Oleg Sviridov","doi":"10.1016/j.ab.2024.115695","DOIUrl":"10.1016/j.ab.2024.115695","url":null,"abstract":"<div><div>Detection of <em>Salmonella</em> in food is topical due to known cases of salmonellosis epidemics. Immunochemical methods including ELISA are widely used for <em>Salmonella</em> detection. Traditionally, commercial ELISA kits are based on sandwich technique and detect lipopolysaccharide (LPS), which is considered to be the component of the outer membrane of Gram-negative bacteria. Our aim was elaboration of competitive ELISA test for <em>Salmonella</em> detection in food with improved parameters. It was shown that in the <em>Salmonella</em> culture after the standard sample preparation procedure LPS is present mainly outside cells as a component of outer membrane vesicles. Improved sample preparation procedure includes separation of bacteria from the medium and analysis of the medium, which increases analytical sensitivity. Immobilization of the bovine serum albumin (BSA)-LPS conjugate in microplate wells allows to obtain a more homogeneous coating than immobilization of LPS itself. Thus, we have developed test system for <em>Salmonella</em> detection in food by competitive ELISA of LPS secreted into the culture medium with the immobilized BSA-LPS conjugate and monoclonal antibodies (mAb) to LPS core in the liquid phase. New competitive ELISA test is high sensitive, give reproducible results, allows the detection of any <em>Salmonella</em> serotype and is important for the protection of human health.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"697 ","pages":"Article 115695"},"PeriodicalIF":2.6,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142493134","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

Micro scale chromatography of human plasma proteins for nano LC-ESI-MS/MS 用于纳米 LC-ESI-MS/MS 的人体血浆蛋白质微尺度色谱法。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-10-22 DOI: 10.1016/j.ab.2024.115694

Zhuo Zhen Chen, Jaimie Dufresne, Peter Bowden, Dominika Celej, Ming Miao, John G. Marshall

{"title":"Micro scale chromatography of human plasma proteins for nano LC-ESI-MS/MS","authors":"Zhuo Zhen Chen, Jaimie Dufresne, Peter Bowden, Dominika Celej, Ming Miao, John G. Marshall","doi":"10.1016/j.ab.2024.115694","DOIUrl":"10.1016/j.ab.2024.115694","url":null,"abstract":"<div><div>Organic precipitation of proteins with acetonitrile demonstrated complete protein recovery and improved chromatography of human plasma proteins. The separation of 25 μL of human plasma into 22 fractions on a QA SAX resin facilitated more effective protein discovery despite the limited sample size. Micro chromatography of plasma proteins over quaternary amine (QA) strong anion exchange (SAX) resins performed best, followed by diethylaminoethyl (DEAE), heparin (HEP), carboxymethyl cellulose (CMC), and propyl sulfate (PS) resins. Two independent statistical methods, Monte Carlo comparison with random MS/MS spectra and the rigorous X!TANDEM goodness of fit algorithm protein p-values corrected to false discovery rate <em>q</em>-values (<em>q</em> ≤ 0.01) agreed on at least 12,000 plasma proteins, each represented by at least three fully tryptic corrected peptide observations. There was qualitative agreement on 9393 protein/gene symbols between the linear quadrupole versus orbital ion trap but also quantitative agreement with a highly significant linear regression relationship between log observation frequency (F value 4,173, p-value 2.2e-16)<strong>.</strong> The use of a QA resin showed nearly perfect replication of all the proteins that were also found using DEAE-, HEP-, CMC-, and <em>P</em>S-based chromatographic methods combined and together estimated the size of the size of the plasma proteome as ≥12,000 gene symbols.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"697 ","pages":"Article 115694"},"PeriodicalIF":2.6,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142493136","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

Uracil base PCR implemented for reliable DNA walking 实施尿嘧啶碱基 PCR，实现可靠的 DNA 走查。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-10-22 DOI: 10.1016/j.ab.2024.115697

Rongrong Wang , Yinwei Gu , Hong Chen , Bingkun Tian , Haixing Li

引用次数: 0

Electrochemical biosensor based on copper sulfide/reduced graphene oxide/glucose oxidase construct for glucose detection 基于硫化铜/还原氧化石墨烯/葡萄糖氧化酶构建的电化学生物传感器用于葡萄糖检测。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-10-22 DOI: 10.1016/j.ab.2024.115696

Yuchen Zhang , Jiangnan Chen , Huifang Wang , Xianghua Gao , Baolong Niu , Wenfeng Li , Hong Wang

{"title":"Electrochemical biosensor based on copper sulfide/reduced graphene oxide/glucose oxidase construct for glucose detection","authors":"Yuchen Zhang , Jiangnan Chen , Huifang Wang , Xianghua Gao , Baolong Niu , Wenfeng Li , Hong Wang","doi":"10.1016/j.ab.2024.115696","DOIUrl":"10.1016/j.ab.2024.115696","url":null,"abstract":"<div><div>Due to the current increase in the number of people suffering from diabetes worldwide, how to monitor the blood glucose level in the human body has become an urgent problem to be solved nowadays. The electrochemical sensor method can be used for real-time glucose monitoring due to its advantages of real-time monitoring capability and high sensitivity. Reduced graphene oxide (rGO) has great potential for application in the field of sensors due to its advantages of large specific surface area, high stability, and good electrical and thermal conductivity. Meanwhile, the synergistic effect between two-dimensional transition metal sulfides and graphene can improve the electrochemical performance of materials due to their similar mechanical flexibility and strength. This article uses flake graphite, copper sulfate, and glucose oxidase (GOx) as raw materials to prepare CuS/rGO/GOx/GCE electrodes, and explores the performance of electrode electrocatalysis for glucose. The results showed that the prepared sensor was characterized by a low detection limit (1.75 nM) and a wide linear range (0.1–100 mM) for glucose detection, displaying a good overall detection performance, and its sensing mechanism and dynamic process were also investigated. In addition, the sensor has outstanding selectivity, anti-interference, repeatability, reproducibility and practicality.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"696 ","pages":"Article 115696"},"PeriodicalIF":2.6,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142493135","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

Impact of Lectin biotinylation for surface plasmon resonance and enzyme-linked Lectin assays for protein glycosylation 表面等离子共振和酶联凝集素检测蛋白质糖基化的凝集素生物素化的影响。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-10-19 DOI: 10.1016/j.ab.2024.115693

Benjamin Serafin , Amine Kamen , Gregory De Crescenzo , Olivier Henry

{"title":"Impact of Lectin biotinylation for surface plasmon resonance and enzyme-linked Lectin assays for protein glycosylation","authors":"Benjamin Serafin , Amine Kamen , Gregory De Crescenzo , Olivier Henry","doi":"10.1016/j.ab.2024.115693","DOIUrl":"10.1016/j.ab.2024.115693","url":null,"abstract":"<div><div>Lectins are widely employed for the assessment of protein glycosylation as their carbohydrate binding specificities have been well characterized. In glycosylation assays, lectins are often conjugated with biotin tags, which interact with streptavidin to functionalize biosensing surfaces or recruit signal generating molecules, depending on the assay configuration. We here demonstrate that a high degree of biotin conjugation can limit total capture to streptavidin functionalized SPR surfaces due to multipoint binding, and can additionally bias the reported kinetic evaluations when measuring the interaction between lectins and glycoproteins by SPR. For microplate assays using different configurations, high biotinylation ratios can effectively amplify the signal obtained when using Streptavidin conjugates for detection, in some cases significantly lowering the limit of detection. The cumulative results express the importance of customizing the ligand biotinylation ratios for different assay configurations, as commercially obtained pre-biotinylated lectins are not necessarily optimized for different assay configurations.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"696 ","pages":"Article 115693"},"PeriodicalIF":2.6,"publicationDate":"2024-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142456264","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

The downstream purification of bispecific antibodies 双特异性抗体的下游纯化。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-10-18 DOI: 10.1016/j.ab.2024.115692

Qian Li , Hongyang Zhao , Xiaoying Liang, Qingquan He, Zicheng Wang, Guohong Qin, GuoZhu Li, Dan Xu

引用次数: 0

Decreased solubility and increased adsorptivity of a biotinylated humanized anti-cocaine mAb 生物素化人源化抗柯卡因 mAb 的溶解度降低，吸附性增强。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-10-18 DOI: 10.1016/j.ab.2024.115690

Terence L. Kirley, Andrew B. Norman

{"title":"Decreased solubility and increased adsorptivity of a biotinylated humanized anti-cocaine mAb","authors":"Terence L. Kirley, Andrew B. Norman","doi":"10.1016/j.ab.2024.115690","DOIUrl":"10.1016/j.ab.2024.115690","url":null,"abstract":"<div><div>Biotinylation of proteins, including antibodies, is a very useful and important modification for a variety of biochemical characterizations, including anti-drug antibody (ADA) assays used to detect antibodies raised against therapeutic antibodies. We assessed different degrees of biotin labeling of an anti-cocaine mAb currently under development for treating cocaine use disorder. We noted that higher levels of biotin labeling dramatically decreased mAb solubility, and increased the tendency to bind to surfaces, complicating characterization of the biotinylated antibody. Specifically, in phosphate buffered saline, labeling stoichiometries of more than about 3 biotin/mAb resulted in decreased recoveries due to increased binding to surfaces and decreased mAb solubility. Native gel agarose electrophoresis, differential scanning fluorimetry, and isothermal titration calorimetry all demonstrated changes in the mAb which became more pronounced above a labeling ratio of 3 biotin/mAb. At 3.0 biotin/mAb, there were minimal changes in solubility, adsorptivity, exposure of hydrophobic dye-binding sites, heat stability, and cocaine binding, in stark contrast to labeling with 5.6 biotin/mAb. Thus, the degree of biotinylation should be kept at about 3 biotin/mAb to maintain antigen binding and general structure, solubility, and stability of this mAb, a finding which may be important for other similar mAbs currently in use or under development.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"696 ","pages":"Article 115690"},"PeriodicalIF":2.6,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142456263","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

Analysis of human milk oligosaccharides from women with gestational diabetes mellitus 分析妊娠糖尿病妇女的母乳低聚糖。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-10-18 DOI: 10.1016/j.ab.2024.115689

Xinyue Ma , Yue Lu , Chuncui Huang , Zhendong Guo , Zheng Xiang , Huanyu Gao , Keli Zhao , Yao Zhao , Yan Li

{"title":"Analysis of human milk oligosaccharides from women with gestational diabetes mellitus","authors":"Xinyue Ma , Yue Lu , Chuncui Huang , Zhendong Guo , Zheng Xiang , Huanyu Gao , Keli Zhao , Yao Zhao , Yan Li","doi":"10.1016/j.ab.2024.115689","DOIUrl":"10.1016/j.ab.2024.115689","url":null,"abstract":"<div><div>Human milk oligosaccharides (HMOs) are bioactive components which play an important role in infant health. HMO composition is vulnerable to changes of maternal conditions including lactation stages and maternal phenotypes. Pregnant diseases such as gestational diabetes mellitus (GDM) are commonly found in women during pragnancy, and may cause disorder in maternal physiological metabolism which is harmful to infants. Unfortunately, anlysis of oligosaccharides from women with GDM is limited. To address this issue, we analyzed HMO compositions and profiles in breast milk from women with GDM using an established 96-well plate permethylation platform and MALDI-TOF-MS. We enrolled 127 women with GDM, and investigated HMO abundances in colostrum, transition milk, and mature milk respectively. We found that GDM affected HMO compositions in breast milk, and the level of fucosylation became higher over the course of lactation for all the women with GDM. Interestingly, the relative abundances of fucosylated HMOs in different lactation stages were affected differentially by GDM, with the most pronounced effect in colostrum. In particular, the relative abundances of H3N1F1 and H3N1F2 sharply decreased over time, showing very low levels in late lactation. These differences in our findings need further investigation to develop optimal feeding for mothers with GDM.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"696 ","pages":"Article 115689"},"PeriodicalIF":2.6,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142456262","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

Chemical composition alterations in rat brain hypothalamus induced by irisin administration using spectroscopic and machine learning techniques 利用光谱学和机器学习技术研究施用鸢尾素诱导的大鼠大脑下丘脑化学成分变化

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-10-16 DOI: 10.1016/j.ab.2024.115687

Zozan Guleken , Huri Dedeakayoğulları , Esra Kutlu , Zeynep Ceylan , Joseph Cebulski , Joanna Depciuch

{"title":"Chemical composition alterations in rat brain hypothalamus induced by irisin administration using spectroscopic and machine learning techniques","authors":"Zozan Guleken , Huri Dedeakayoğulları , Esra Kutlu , Zeynep Ceylan , Joseph Cebulski , Joanna Depciuch","doi":"10.1016/j.ab.2024.115687","DOIUrl":"10.1016/j.ab.2024.115687","url":null,"abstract":"<div><div>This study employed Fourier transform infrared (FTIR) spectroscopy to determine the chemical composition of brain tissues and the changes induced by irisin at doses of 50 mg and 100 mg. Brain tissues were collected from control rats and those administered with irisin, and key vibrational peaks were analyzed. In the 50 mg irisin group, all described vibrations decreased compared to control tissues, while the 100 mg group showed a decrease only in lipid vibrations. Comparatively, the 50 mg group had lower absorbance of phospholipids, amides, and lipid functional groups than the 100 mg group. Lower amounts of these compounds were found in treated tissues compared to controls, with higher levels in the 100 mg group. Ratios between amide peaks revealed significant differences between groups. Principal component analysis (PCA) differentiated control and irisin-treated tissues, primarily using PC1 and PC3. The decision tree model exhibited high classification accuracy, especially in the 800–1800 cm⁻<sup>1</sup> range, with high sensitivity and specificity. FTIR spectroscopy effectively highlighted chemical changes in brain tissues due to irisin, demonstrating dose-dependent variations. The combination of PCA, ROC analysis, and decision tree modeling underscored the potential of FTIR spectroscopy for studying the biochemical effects of compounds like irisin.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"696 ","pages":"Article 115687"},"PeriodicalIF":2.6,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142445775","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

A novel simple fluorometric protease assay for monitoring hydrolysis of proteins in real time 用于实时监测蛋白质水解的新型简易荧光蛋白酶测定法

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-10-16 DOI: 10.1016/j.ab.2024.115688

Miha Bahun , Nataša Poklar Ulrih

引用次数: 0