Analytical biochemistry最新文献_第2页

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":null,"pages":null},"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":null,"pages":null},"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":null,"pages":null},"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":null,"pages":null},"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

Photoelectrochemical biosensors: Prospects of graphite carbon nitride-based sensors in prostate-specific antigen diagnosis 光电化学生物传感器：基于氮化石墨的传感器在前列腺特异性抗原诊断中的应用前景。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-10-10 DOI: 10.1016/j.ab.2024.115686

Seyed Saman Nemati, Gholamreza Dehghan

{"title":"Photoelectrochemical biosensors: Prospects of graphite carbon nitride-based sensors in prostate-specific antigen diagnosis","authors":"Seyed Saman Nemati, Gholamreza Dehghan","doi":"10.1016/j.ab.2024.115686","DOIUrl":"10.1016/j.ab.2024.115686","url":null,"abstract":"<div><div>Prostate cancer (PC) is very common in old age and causes many deaths. Early diagnosis and monitoring of the progress of the disease and the effectiveness of PC treatment are critical. On the other hand, choosing a specific biomarker for PCs is essential. Prostate-specific antigen (PSA) is a specific biomarker secreted in the prostate epithelial cells, which increases in cancer cells. Between all employed sensing mechanism, electrochemical sensors based on nanomaterials have created many hopes. Meanwhile, graphite carbon nitride (g-C<sub>3</sub>N<sub>4</sub>) is interested in developing photoelectrochemical sensors due to its large surface area, stability, easy modification, and good photoelectronic properties. In this review, electrochemical sensors based on nanocomposites containing g-C<sub>3</sub>N<sub>4</sub> have been investigated in PSA detection. After providing an overview of the characteristics of g-C<sub>3</sub>N<sub>4</sub> and cancer biomarkers, it reviews the strategies and mechanisms involved in identifying PSA. Different approaches to photoelectrochemistry, impedimetric immunosensors, photocatalysis, and luminescence have been used in diagnostic mechanisms. Then, challenges and prospects for electrochemical sensors based on nanocomposites containing g-C<sub>3</sub>N<sub>4</sub> in PSA detection have been analyzed. The recent review generally opens an efficient view in PSA diagnosis and the application of g–C<sub>3</sub>N<sub>4</sub>–based electrochemical sensors in personalized medicine diagnosis and treatment.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142405938","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

Urea biosensors based in zeolites and chalcogenide-oxide semiconductor thin films as active materials: A review 以沸石和氧化镓半导体薄膜为活性材料的尿素生物传感器：综述。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-10-05 DOI: 10.1016/j.ab.2024.115685

Manuel A.Chairez Ortega , Rafael E. González Landaeta , Francisco S. Aguirre Tostado , Soledad V. Torres Argüelles , Amanda Carrillo Castillo

{"title":"Urea biosensors based in zeolites and chalcogenide-oxide semiconductor thin films as active materials: A review","authors":"Manuel A.Chairez Ortega , Rafael E. González Landaeta , Francisco S. Aguirre Tostado , Soledad V. Torres Argüelles , Amanda Carrillo Castillo","doi":"10.1016/j.ab.2024.115685","DOIUrl":"10.1016/j.ab.2024.115685","url":null,"abstract":"<div><div>Diagnosis of renal failure by measuring urea levels has been a topic of intense study in recent years. A major focus has been on improving the sensitivity, linearity, precision, accuracy, and selectivity of biosensors for measuring urea. Although various materials have been used in the fabrication of urea biosensors, ceramics, and chalcogenides have been less explored in this field. Recently, the use of ceramics such as zeolite has been investigated to improve enzyme immobilization methods in urea biosensors and their application in ion-selective membranes, to increase the specificity of the devices. While oxides have been widely used as transducers in urea biosensors, chalcogenide semiconductor materials from Group VI of the periodic table also show promising properties, such as chemical stability, to signal transduction capability, and improved electrical measurements. This review provides a comprehensive overview of recent research in urea biosensors, with a special emphasis on the use of ceramics for enzyme immobilization and chalcogenides as transducers and how these materials contribute to improving the performance of these devices.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-10-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142387379","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

The method of capillary electrophoresis for quantitative determination of hydrophobized hyaluronic acid in its micellar forms 毛细管电泳法，用于定量测定胶束形式的疏水性透明质酸。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-10-04 DOI: 10.1016/j.ab.2024.115684

Yu Ermolenko, M. Antonova, A. Polivanova, S. Gelperina

{"title":"The method of capillary electrophoresis for quantitative determination of hydrophobized hyaluronic acid in its micellar forms","authors":"Yu Ermolenko, M. Antonova, A. Polivanova, S. Gelperina","doi":"10.1016/j.ab.2024.115684","DOIUrl":"10.1016/j.ab.2024.115684","url":null,"abstract":"<div><div>Micelles based on hydrophobized hyaluronic acid (HA) are frequently used in targeted drug delivery systems. Capillary zone electrophoresis (CZE) was utilized for the quantitative determination of hydrophobized and native HA. A universal methodology was developed, suitable for the quantitative analysis of amphiphilic derivatives of hyaluronan (oleyl hyaluronan and hyaluronan conjugate with naphthalimide fluorophore) and native HA with varying molecular weights (15, 150, and 800 kDa). Furthermore, methodologies were proposed for the simultaneous quantification of a drug substance and oleyl hyaluronan in micellar forms based on the latter. The CE technique was applied for analyzing oleyl-hyaluronan-based micellar forms of two poorly soluble drug substances with oppositely charged ionic forms (loperamide and rifabutin). The examples contained in the study demonstrate a range of analytical sensitivity (LOD) for hyaluronan from 11 to 40 μg/mL and for the drug substance from 0.4 to 0.6 μg/mL. The study also showcases the accurate quantitative determination of rifabutin and loperamide in oleyl-hyaluronan-based micellar forms without the need for sample preparation. Thus, the proposed methodologies can be used to quantify native HA or its amphiphilic derivatives and simultaneously determine drug substances of various nature.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142379024","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 time-saving one-step polyacrylamide gel with a colored stacking gel for SDS-PAGE and western blotting 一步式聚丙烯酰胺凝胶，带彩色叠层凝胶，用于 SDS-PAGE 和 Western 印迹，省时省力。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-09-26 DOI: 10.1016/j.ab.2024.115680

Makoto Hagiwara

引用次数: 0