Analytical biochemistry最新文献_第6页

Survey of chemical unfolding complexity as a unique stability assessment assay for monoclonal antibodies 化学展开复杂性作为单克隆抗体独特稳定性评估方法的研究进展。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-11-29 DOI: 10.1016/j.ab.2024.115729

J. Alaina Floyd , Jeremy M. Shaver

{"title":"Survey of chemical unfolding complexity as a unique stability assessment assay for monoclonal antibodies","authors":"J. Alaina Floyd , Jeremy M. Shaver","doi":"10.1016/j.ab.2024.115729","DOIUrl":"10.1016/j.ab.2024.115729","url":null,"abstract":"<div><div>Seventy-two intentionally sequence-diverse antibody variable regions were selected, expressed as IgG1 antibodies, and evaluated by chemical unfolding to survey the complexities of denaturant induced unfolding behavior. A two-transition fit well described the curves and uncovered a wide range of sensitivities to denaturant. Four general types of unfolding curves were observed: balanced traces (each transition responsible for half of the total unfolding curve), low-unfolding traces (first transition is a majority of the unfolding curve), high-unfolding traces (the second transition is the majority of the unfolding curve), and coincident traces (the two transitions are found close to each other, approximating a single transition). The complexity of the data from this survey indicates that focusing on the first inflection point or fitting a single transition model is likely an over-simplistic method for measuring stability by the chemical unfolding assay. Additionally, other conformational assays, such as thermal and low pH unfolding, showed no correlation with the chemical unfolding results, indicating that each of these assays provide alternate information on the different pathways of antibody conformational stability. These results provide a basis for beginning to better connect unfolding behavior to other physical phenotypic behaviors and production process behaviors.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"698 ","pages":"Article 115729"},"PeriodicalIF":2.6,"publicationDate":"2024-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142765576","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 sensing of dopamine using nanostructured silver chromate: Development of an IoT-integrated sensor 纳米结构铬酸银对多巴胺的电化学传感：物联网集成传感器的开发。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-11-29 DOI: 10.1016/j.ab.2024.115726

Lungelo Mgenge , Chandan Saha , Pooja Kumari , Sarit K. Ghosh , Harishchandra Singh , Kaushik Mallick

{"title":"Electrochemical sensing of dopamine using nanostructured silver chromate: Development of an IoT-integrated sensor","authors":"Lungelo Mgenge , Chandan Saha , Pooja Kumari , Sarit K. Ghosh , Harishchandra Singh , Kaushik Mallick","doi":"10.1016/j.ab.2024.115726","DOIUrl":"10.1016/j.ab.2024.115726","url":null,"abstract":"<div><div>Dopamine, one of the most important neurotransmitters, plays a crucial role in the functions of human metabolism, as well as the cardiovascular, central nervous and hormonal systems. This study focuses on the synthesis of nanostructured silver chromate and their application in dopamine sensing. The nanoparticles were synthesized using a complexation-mediated route using aminosalicylic acid as a stabilizer, resulting in uniform particles ranging from 3 to 15 nm in size. The synthesized silver chromate was characterized using X-ray diffraction, transmission electron microscopy and X-ray photoelectron spectroscopy techniques. Electrochemical studies revealed that the silver chromate exhibit excellent catalytic activity for the detection of dopamine. The electroanalysis provided the selective recognition of dopamine with the limit of detection of 1.05 μM and sensitivity of 2.68 μA μM<sup>−1</sup> cm<sup>−2</sup> in a linear range of 5–45 μM. Additionally, a portable, IoT (internet of things)-integrated sensor based on the synthesized silver chromate was developed using Arduino Uno R4 Wi-Fi module, enabling real-time monitoring of dopamine with data transmission to a cloud platform.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"698 ","pages":"Article 115726"},"PeriodicalIF":2.6,"publicationDate":"2024-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142765573","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

Lateral flow immunoassay based on aggregation induced emission nanobeads for the sensitive and accurate detection of chloramphenicol in pig hair 基于聚集诱导发射纳米微球的横向流动免疫分析法灵敏、准确地检测猪毛中氯霉素。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-11-27 DOI: 10.1016/j.ab.2024.115728

Keyang Lai , Jun Xu , Kai Luo , Min Xie , Yuan Chen , Fan Li , Yaomin Zhou , Lihui Gong , Yonghua Xiong , Weihua Lai

{"title":"Lateral flow immunoassay based on aggregation induced emission nanobeads for the sensitive and accurate detection of chloramphenicol in pig hair","authors":"Keyang Lai , Jun Xu , Kai Luo , Min Xie , Yuan Chen , Fan Li , Yaomin Zhou , Lihui Gong , Yonghua Xiong , Weihua Lai","doi":"10.1016/j.ab.2024.115728","DOIUrl":"10.1016/j.ab.2024.115728","url":null,"abstract":"<div><div>Chloramphenicol (CAP) is a once widely used antibiotic, which is able to cause great harm to human health, is banned in some countries or organizations such as China, USA and the European Union for animal breeding. Because CAP in pig hair degraded slower and had amount of residues, pig hair could be used as the target to detect CAP residues. In this study, a competitive lateral flow immunoassay (LFIA), whose label was aggregation induced emission fluorescent nanobeads (AIEFN), was firstly developed for the detection of CAP in pig hair samples. It exhibited a low limit of detection (0.001 μg/kg) and a broad linear range (0.0025–0.32 μg/kg). The recovery rate and coefficient variation with spiked pig hair samples were 88.50%–106.17 % and 1.01%–7.37 %, which indicated good accuracy of this assay. The result of AIEFN-LFIA for the detection of CAP in pig hair was consistent with that of liquid chromatography-mass spectrometry. This assay was rapid with the total detection duration of about 20 min. AIEFN-LFIA was able to be used for rapid, sensitive, accurate and convenient detection of CAP residues in pig hair samples.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"698 ","pages":"Article 115728"},"PeriodicalIF":2.6,"publicationDate":"2024-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142749603","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 sandwich ELISA for the quantification of the anticancer peptide CIGB-552 in human plasma 夹心ELISA法定量测定人血浆中抗癌肽CIGB-552。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-11-27 DOI: 10.1016/j.ab.2024.115725

Nivaldo Angel Gómez Hernández , Gilda Lemos Pérez , Amalia Vazquez Arteaga , Hilda Elisa Garay Pérez , Brizaida Oliva Arguellez , Ania Cabrales Rico , Airela Llamo Guardia , Julio Raúl Fernández Massó

{"title":"A sandwich ELISA for the quantification of the anticancer peptide CIGB-552 in human plasma","authors":"Nivaldo Angel Gómez Hernández , Gilda Lemos Pérez , Amalia Vazquez Arteaga , Hilda Elisa Garay Pérez , Brizaida Oliva Arguellez , Ania Cabrales Rico , Airela Llamo Guardia , Julio Raúl Fernández Massó","doi":"10.1016/j.ab.2024.115725","DOIUrl":"10.1016/j.ab.2024.115725","url":null,"abstract":"<div><div>CIGB-552 is a synthetic anticancer peptide that has been evaluated in vitro and in vivo in lung and colon cancer models. To optimize therapy in the clinic, pharmacokinetic studies are necessary. Previously, a sandwich-type enzyme-linked immunosorbent assay (ELISA) had been developed by our working group for the quantification of CIGB-552 in biological matrices. The objective of this work was to carry out the full validation of the ELISA to support its application in clinical trials. First, we obtained a polyclonal antibody specific for CIGB-552 and with purity greater than 95 %. The lower limit of quantification and the upper limit of quantification were 3125 ng/ml and 200 ng/ml, respectively. The method is exact and precise in the quantification of the peptide with relative error and coefficient of variation values less than 20 %. The ELISA is specific in the presence of CIGB-552 metabolites in the sample, and also presents robustness to certain protocol variations. In summary, the validated ELISA meets the requirements for its application in upcoming clinical trials as part of pharmacokinetic studies.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"698 ","pages":"Article 115725"},"PeriodicalIF":2.6,"publicationDate":"2024-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142749497","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

High-throughput sperm screening using one-step RT-qPCR: Improvement and re-evaluation 一步RT-qPCR高通量精子筛查：改进和重新评估。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-11-27 DOI: 10.1016/j.ab.2024.115727

Seiji Kubo , Keito Amai , Fumitaka Nakano , Jin Tanaka , Hideki Niimi

引用次数: 0

Comparative validation of UV-spectrophotometry and RP-HPLC methods for cefixime and moxifloxacin analysis 紫外分光光度法和 RP-HPLC 法用于头孢克肟和莫西沙星分析的比较验证

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-11-26 DOI: 10.1016/j.ab.2024.115724

Mahesh Chaudhari , Parul K. Parmar , Kiran Dudhat

{"title":"Comparative validation of UV-spectrophotometry and RP-HPLC methods for cefixime and moxifloxacin analysis","authors":"Mahesh Chaudhari , Parul K. Parmar , Kiran Dudhat","doi":"10.1016/j.ab.2024.115724","DOIUrl":"10.1016/j.ab.2024.115724","url":null,"abstract":"<div><h3>Aim</h3><div>This study presents the development and validation of UV-spectrophotometry and RP-HPLC methods for the simultaneous quantification of Cefixime Trihydrate (CEFI) and Moxifloxacin Hydrochloride (MOXI) in pharmaceutical formulations.</div></div><div><h3>Methodology</h3><div>Two UV-spectrophotometric methods, including the absorbance ratio (Q-Absorption) and First Order Derivative Spectroscopy, were developed and validated for their linearity, precision, accuracy, and sensitivity. Additionally, a robust RP-HPLC method using a C18 column and optimized mobile phase was employed for efficient separation and simultaneous estimation of CEFI and MOXI. All methods were validated in accordance with ICH guidelines, with system suitability parameters confirming the reliability of the RP-HPLC method for routine analysis.</div></div><div><h3>Results</h3><div>The absorbance ratio and First Order Derivative methods demonstrated low %R.S.D values, high accuracy, and satisfactory sensitivity for both drugs. Similarly, the RP-HPLC method achieved high resolution, precision, and robustness. Statistical analysis through ANOVA revealed no significant differences between the methods in terms of accuracy and precision. The methods were applied to analyze marketed formulations, further confirming their applicability in routine quality control.</div></div><div><h3>Conclusion</h3><div>In conclusion, the validated methods provide accurate, precise, and sensitive techniques for the simultaneous estimation of CEFI and MOXI, making them suitable for pharmaceutical quality control and regulatory compliance.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"697 ","pages":"Article 115724"},"PeriodicalIF":2.6,"publicationDate":"2024-11-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142738136","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 assay for measuring acetyl-coenzyme a carboxylase activity in grasses using malachite green 利用孔雀石绿验证测定草中乙酰辅酶 A 羧化酶活性的方法

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-11-23 DOI: 10.1016/j.ab.2024.115723

Yoshinobu Jin

{"title":"Validation of assay for measuring acetyl-coenzyme a carboxylase activity in grasses using malachite green","authors":"Yoshinobu Jin","doi":"10.1016/j.ab.2024.115723","DOIUrl":"10.1016/j.ab.2024.115723","url":null,"abstract":"<div><div>Acetyl-CoA carboxylase (ACCase) is one of the most important enzymes as a herbicide target in gramineous plant species, however, assay methods for the enzyme are primarily limited to those using radioisotopes (RI). Typically, the measurement method that uses RI necessitates specialized facilities and equipment, and involves complex procedures throughout the experiment. As another method for detecting ACCase activity, the colorimetric method using malachite green (MG) is also known. However, reports on this method are limited, and information regarding the simplicity of the procedure and the scope of its application remains unclear. To better understand the method using MG and to develop a simpler assay method, crude enzymes extracted from various target-site resistant (TSR) biotypes of blackgrass (<em>Alopecurus myosuroides</em>) were examined in enzyme inhibition tests. As a result, this method was able to accurately detect the relationship between the chemical classes of ACC inhibitors and cross-resistance to specific TSRs. Moreover, the ACCase activity of other grass species was also examined using this method. By using crude enzymes from various species and a commercially available phosphatase kit containing MG, ACCase activity was detectable with good accuracy. In addition, enzyme inhibition studies using ACCase inhibiting herbicides revealed that this method reproduced results similar to those obtained with the RI method. The Z′-factor, an index of high-throughput screening (HTS), was around 0.7, indicating that it is an excellent screening system. These results suggest that the assay method using MG is very simple, labor-saving, and accurate with a throughput much higher than that of the existing RI method. Therefore, it is strongly suggested that the method could replace the RI method in most cases. These results indicate that it is applicable to HTS for ACCase inhibitors.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"697 ","pages":"Article 115723"},"PeriodicalIF":2.6,"publicationDate":"2024-11-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142715228","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

In vitro selection of dye-fluorescence-enhancing peptide aptamer by cDNA display 通过 cDNA 展示体外选择染料荧光增强多肽适配体。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-11-23 DOI: 10.1016/j.ab.2024.115722

Takashi Kubo , Tomoyuki Koike , Tomoki Ouchi , Nayab Khaliq , Eita Sasaki , Kouichi Kuroda , Mitsuyoshi Ueda , Kenjiro Hanaoka , Naoto Nemoto

引用次数: 0

Sample loading in gel electrophoresis using adapted 3D printers 使用适配的 3D 打印机在凝胶电泳中装载样品。

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-11-23 DOI: 10.1016/j.ab.2024.115721

Tao Tao , Alexey Kopyl , Yuki Yew , Fawaz El-Dani , Hassan Ali Abid , Thomas Hiscox , Oi Wah Liew , Tuck Wah Ng

引用次数: 0

Studying C9orf72 dipeptide repeat polypeptide aggregation using an analytical ultracentrifuge equipped with fluorescence detection 利用配备荧光检测装置的分析型超速离心机研究 C9orf72 二肽重复多肽的聚集情况

IF 2.6 4区生物学

Analytical biochemistry Pub Date : 2024-11-22 DOI: 10.1016/j.ab.2024.115720

Bashkim Kokona , Nicole R. Cunningham , Jeanne M. Quinn , Danielle R. Jacobsen , F. Jay Garcia , Sierra M. Galindo , Leonard Petrucelli , Walter F. Stafford , Thomas M. Laue , Robert Fairman

{"title":"Studying C9orf72 dipeptide repeat polypeptide aggregation using an analytical ultracentrifuge equipped with fluorescence detection","authors":"Bashkim Kokona , Nicole R. Cunningham , Jeanne M. Quinn , Danielle R. Jacobsen , F. Jay Garcia , Sierra M. Galindo , Leonard Petrucelli , Walter F. Stafford , Thomas M. Laue , Robert Fairman","doi":"10.1016/j.ab.2024.115720","DOIUrl":"10.1016/j.ab.2024.115720","url":null,"abstract":"<div><div>Sedimentation velocity, using an analytical ultracentrifuge equipped with fluorescence detection, and electrophoresis methods are used to study aggregation of proteins in transgenic animal model systems. Our previous work validated the power of this approach in an analysis of mutant huntingtin aggregation<em>.</em> We demonstrate that this method can be applied to another neurodegenerative disease studying the aggregation of three dipeptide repeats (DPRs) produced by aberrant translation of mutant <em>c9orf72</em> containing large G<sub>4</sub>C<sub>2</sub> hexanucleotide repeats<em>.</em> These repeat expansions are the most common cause of familial amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). We analyzed the aggregation patterns of (Gly-Pro)<sub>47</sub>, (Gly-Ala)<sub>50</sub>, and (Gly-Arg)<sub>50</sub> fused to fluorescent proteins in samples prepared from <em>D. melanogaster</em>, and (Gly-Ala)<sub>50</sub> in <em>C. elegans</em>, using AU-FDS and SDD-AGE<em>.</em> Results suggest that (GP)<sub>47</sub> is largely monomeric. In contrast, (GA)<sub>50</sub> forms both intermediate and large-scale aggregates. (GR)<sub>50</sub> is partially monomeric with some aggregation noted in SDD-AGE analysis. The aggregation of this DPR is likely to represent co-aggregated states with DNA and/or RNA. The power of these methods is the ability to gather data on aggregation patterns and characteristics in animal model systems, which may then be used to interpret the mitigation of aggregation through genetic or molecular therapeutic interventions.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"697 ","pages":"Article 115720"},"PeriodicalIF":2.6,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142705120","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