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Hapten design to prepare monoclonal antibodies and establishment of immunoassay for direct screening of oxamichydrazide in chicken,the metabolite of nifuraldezone

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One Health Advances 2023年第1期1卷 311-322页

作者： Ghulam Mujtaba Mari Weilin Wu Aisha Talpur Jan Muhammad Shah Saqib Ali Faiz Khand Erum Bughio Abdul Samad Zhanhui Wang sergei a.eremin Department of Veterinary Pharmacology and Toxicology Faculty of Bio-SciencesShaheed Benazir Bhutto University of Veterinary and Animal SciencesSakrand 67210Pakistan National Key Laboratory of Veterinary Public Health Security Beijing Key Laboratory of Detection Technology for Animal-Derived Food SafetyCollege of Veterinary MedicineChina Agricultural UniversityBeijing 100193China Department of Veterinary Surgery and Obstructers Faculty of Veterinary SciencesShaheed Benazir Bhutto University of Veterinary and Animal SciencesSakrand 67210Pakistan Department of Poultry Production Faculty of Animal Production and TechnologyShaheed Benazir Bhutto University of Veterinary and Animal SciencesSakrand 67210Pakistan Department of Dairy Technology Faculty of Animal Production and TechnologyShaheed Benazir Bhutto University of Veterinary and Animal SciencesSakrand 67210Pakistan Department of Chemistry M.V.Lomonosov Moscow State University119991 MoscowRussia

In this study,two novel haptens of oxamichydrazide(OXZ),the metabolite of nifuraldezone,were designed and synthesized by derivatization with ethyl-4-bromobutanoate and ethyl-2-bromoacetate for the first *** rationality of the haptens was enlightened by conformational alignment and electronic evaluations based on computional *** synthesized haptens were efficient in generating antibody response to OXZ.A monoclonal antibody(mAb)named 2D9 with high affinity was obtained and used to establish an indirect competitive enzyme-linked immunosorbent assay(icELISA)for the direct determination of OXZ in chicken samples without the need for derivatization *** optimum conditions,the developed icELISA showed an IC50 value of 0.66 ng mL^(−1),and limit of detection of 0.11μg kg^(−1)in *** average recoveries were found to be 80.4-91.3%with a coefficient of variation less than 9.0%.In conclusion,we have successfully desigened haptens,prepared mAbs and developed an immunoassay for the direct determination of OXZ in chicken samples without the need for a derivatization step for the first *** provides a potentially rapid,accurate,and sensitive screening tool for nifuraldezone residue in animal-derived food.

关键词： Nifuraldezone Oxamichydrazide Hapten design Monoclonal antibody icELISA Direct determination

Chemiluminescence resonance energy transfercompetitive immunoassay employing haptenfunctionalizedquantum dots for the detection of a chemical contaminant

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Chemiluminescence resonance energy transfercompetitive immun...

荧光偏振免疫分析在农药和兽药残留检测中的研究进展

第七届中国畜牧科技论坛

作者： Ma mingfang Wen kai Ross C.Beier sergei a.eremin Li chenglong Zhang suxia Wang zhanhui Shen jianzhong

We describe here a new strategy for the use of chemiluminescence resonance energy transfer(CRET)by usinghapten-functionalized quantum dots(QDs)as the acceptor in a competitive immunoassay for the detection of a chemic...

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光谱学与光谱分析 2007年第11期27卷 2299-2306页

作者：王战辉张素霞沈建忠 sergei a.eremin 中国农业大学动物医学院北京100094 莫斯科大学化学院莫斯科119992

荧光偏振免疫分析技术是以免疫竞争原理和荧光偏振原理为基础的一种以快速筛选为目的的分析技术,主要用于测定小分子量物质(抗原),就是检测荧光标记抗原在结合特异性的抗体前后,荧光偏振值的变化,是均相的竞争免疫分析方法,反应系统完... 详细信息

荧光偏振免疫分析技术是以免疫竞争原理和荧光偏振原理为基础的一种以快速筛选为目的的分析技术,主要用于测定小分子量物质(抗原),就是检测荧光标记抗原在结合特异性的抗体前后,荧光偏振值的变化,是均相的竞争免疫分析方法,反应系统完全在溶液中,不需要分离结合的和未结合的抗体,不受溶液颜色、仪器灵敏度变化的影响,该技术最显著的特点是操作简单和检测时间短,适用于大量样本的快速筛选检测。目前该技术在农药和兽药残留分析中已经得到了应用,但是国内的相关研究尚未见报道。文章综述了荧光偏振免疫分析的历史背景,发展历程和主要原理,概括了荧光偏振免疫分析方法在农药和兽药残留分析中的研究进展。

关键词：荧光偏振免疫分析测定农药和兽药

Enzyme-Linked Immunosorbent Assay and Immunochromatographic Strip for Rapid Detection of Atrazine in Three Medicinal Herbal Roots

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World Journal of Traditional Chinese Medicine 2021年第1期7卷 97-103页

作者： Yu-Dan Wang Jia-An Qin Liu Wu Bao-Min Wang sergei eremin Shi-Hai Yang Mei-Hua Yang Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine Ministry of EducationInstitute of Medicinal Plant DevelopmentChinese Academy of Medical SciencesPeking Union Medical CollegeBeijingChina College of Traditional Chinese Medicine Jilin Agricultural UniversityChangchunChina College of Agronomy and Biotechnology China Agricultural UniversityBeijingChina Department of Chemical Enzymology Faculty of Chemistry M.V.Lomonosov Moscow State University Leninsky Gory1119991 MoscowRussia

Objectives:An enzyme-linked immunosorbent assay(ELISA)and colloidal gold-based immunochromatographic(ICG)strip assay will be developed for the rapid and high-throughput detection of atrazine(ATZ)in medicinal ***:A monoclonal antibody against ATZ was obtained after the immunization of mice,cell fusion,and hybridoma screening,and the antibody was used to develop direct competitive ELISA(dcELISA)and the ICG strip ***:Both dcELISA and ICG strip methods were established,optimized,and validated for the detection of ATZ in Salviae miltiorrhizae radix et rhizome,Astragali radix,and Isatidis *** had a half-maximum inhibition concentration of 10.56 ng/m L(Salviae miltiorrhizae radix et rhizome),7.6 ng/m L(Astragali radix),and 8.15 ng/m L(Isatidis radix).The limit of detection(LOD)of the ICG strip assay was 12.5 ng/mL(Salviae miltiorrhizae radix et rhizome),12.5 ng/mL(Astragali radix),and 6.25 ng/mL(Isatidis radix)in different herb *** to the recognition characteristics of the monoclonal antibody for the pesticides ATZ,propazine,sebuthylazine,and prometryn,the detection results of real samples by the two immunoassays were confirmed by liquid chromatography–tandem mass spectrometry,which proved the accuracy and reliability of the established ***:The proposed dcELISA and ICG strip methods were suitable for the rapid,convenient,and high-throughput detection of ATZ in these medicinal herbs.

关键词： Astragali radix atrazine isatidis radix monoclonal antibody salviae miltiorrhizae radix et rhizome

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荧光偏振免疫分析方法快速检测沙拉沙星残留

分析化学 2012年第8期40卷 1247-1251页

作者：宋佩孟萌 sergei a.eremin 张太昌田溪薛虎寅张昱尹永梅郗日沫南开大学药学院南开大学药物化学生物学国家重点实验室天津市分子药物研究重点实验室 Faculty of Chemistry M.V.Lomonosov Moscow State University

以异硫氰酸荧光素(FITC)标记沙拉沙星合成荧光标记物,采用薄层色谱法提纯,优化了反应时间、标记物和抗体的工作浓度,建立了沙拉沙星的快速荧光偏振免疫分析法(FPIA)。本方法测定沙拉沙星在缓冲液中的半数抑制浓度(IC50)为43.2μg/L;检... 详细信息

以异硫氰酸荧光素(FITC)标记沙拉沙星合成荧光标记物,采用薄层色谱法提纯,优化了反应时间、标记物和抗体的工作浓度,建立了沙拉沙星的快速荧光偏振免疫分析法(FPIA)。本方法测定沙拉沙星在缓冲液中的半数抑制浓度(IC50)为43.2μg/L;检测范围为5.7～327μg/L,可以达到国家规定的动物性食品中兽药最高残留限量(80μg/kg)的要求。本研究考察了FPIA测定沙拉沙星的动力学过程及对其它4种喹诺酮类药物的交叉反应。结果表明,环丙沙星、恩诺沙星、加替沙星及氧氟沙星的交叉反应率分别为3.3%,1.8%,1.7%和0.7%。在牛奶和猪尿中沙拉沙星的回收率分别在71%～94%和74%～102%之间。本方法操作简单快捷,整个检测过程只需5 min、而且灵敏度较高、特异性强,适用于动物性食品中沙拉沙星残留的快速筛选检测。

关键词：荧光偏振免疫分析沙拉沙星喹诺酮类药物荧光标记

The Development of A Fluorescence Polarization Immunoassay for Aflatoxin Detection

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Biomedical and Environmental Sciences 2014年第2期27卷 126-129页

作者： SHENG Ya Jie eremin sergei MI Tie Jun ZHANG Su Xia SHEN Jian Zhong WANG Zhan Hui Department of Veterinary Pharmacology and Toxicology College of Veterinary Medicine China Agricultural University and Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety Beijing Laboratory for Food Quality and Safety Department of Chemistry M.V. Lomonosov Moscow State University

A fluorescence polarization immunoassay （FPIA） was developed for the analysis ofaflatoxins （AFs） using an anti-aflatoxin B1 （AFB1） monoclonal antibody and a novel fluorescein-labeled AFB1 tracer. The FPIA showed an IC50 value of 23.33 ng/mL with a limit of detection of 13.12 ng/mL for AFB1. The cross-reactivities of AFB1, AFB2, AFG1, AFG2, AFM1, and AFM2 with the antibody were 100%, 65.7%, 143%, 23.5%, 111.4%, and 2%, respectively. The group-specificity of anti-AFB1mAb indicated that the FPIA could potentially be used in a screening method for the detection of total AFs, albeit not AFG2 and AFM2. The total time required for analyzing 96 samples in one microplate was less than 5 rain. This study demonstrates the potential usefulness of the FPIA as a rapid and simple technique for monitoring AFs.

关键词： FPIA AFB The Development of A Fluorescence Polarization Immunoassay for Aflatoxin Detection AFM EDF

黄曲霉素M1间接竞争ELISA的建立及其在牛奶中的应用

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科学技术与工程 2013年第19期21卷 5455-5458页

作者：陈曦孟萌 sergei a.eremin 许迎辉尹永梅郗日沫天津科技大学生物工程学院工业发酵微生物教育部重点实验室天津市工业微生物重点实验室天津300457 南开大学药学院南开大学药物化学生物学国家重点实验室天津市分子药物研究重点实验室天津300071 莫斯科大学化学学院莫斯科119991 河北省石家庄市餐饮化保执法大队石家庄050000

利用高特异性单克隆抗体,建立黄曲霉素M1的间接竞争ELISA检测方法。棋盘法优化了包被抗原及酶标抗体的最佳稀释倍数。包被抗原和酶标抗体的最佳工作浓度分别为1∶15000和1∶2000。该方法的线性范围为(0.1—8.1)ng·mL-1。除了与黄曲霉... 详细信息

利用高特异性单克隆抗体,建立黄曲霉素M1的间接竞争ELISA检测方法。棋盘法优化了包被抗原及酶标抗体的最佳稀释倍数。包被抗原和酶标抗体的最佳工作浓度分别为1∶15000和1∶2000。该方法的线性范围为(0.1—8.1)ng·mL-1。除了与黄曲霉素B1的交叉反应为35%外,与黄曲霉素B2、黄曲霉素G1及黄曲霉素G2的交叉反应均小于1%。在牛奶实际样品检测的回收率均大于70%,而且检测过程在10min以内即可完成。该方法灵敏度高,特异性强,而且操作简单,适合多样本的快速筛查,为黄曲霉素M1的高效检测提供了一个良好的选择。

关键词：黄曲霉素M1 间接竞争ELISA 交叉反应牛奶

Antibody Production for a Rapid Fluorescence Polarization Immunoassay of Estrone

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Biomedical and Environmental Sciences 2014年第1期27卷 52-55页

作者： ZHANG Xuan WANG Qiang YU Zhong sergei A. eremin YU Chun Fai LIU Jin SUN Yuan Ming LEI Hong Tao Risk Assessment Key Laboratory of Agricultural Product in Preserving Ministry of Agriculture/Guangdong Provincial Key Laboratory of Food Quality and SafetySouth China Agricultural University Department of Chemistry M.V. Lomonosov Moscow State University Environmental Science Program United International College

Estrone has been identified as a potential endocrine-disrupting chemical （EDC）[1]. Estrone is usually quantified by gas chromatography-mass spectrometry （GC-MS）, GC-MS/MS, high performance liquid chromatography （HPLC）, HPLC- MS, and HPLC-MS/MS, etc.[2-3]. Meanwhile, several immunoassays based on radioimmunoassay, enzyme linked immunosorbent assay （ELISA） or chemiluminescence immunoassay （CLIA） for determination of estrone in real samples have been developed[2＇4]. Although these methods are sensitive, they need multistage separation and are thus time-consuming and laborious. A very promising way for the simplification of immunoassays for routine applications is a shift from heterogeneous methods （with separation） to homogeneous assays （without separation）[5]. Fluorescence polarization immunoassay （FPIA） is one of the homogeneous techniques that meets the requirements of a simple, reliable, fast, and cost-effective analysis[6]. Therefore, the present study is focused on the development of FPIA in order to analyze estrone based on antibody production.

关键词： FPIA Antibody Production for a Rapid Fluorescence Polarization Immunoassay of Estrone

手性药物与生物大分子相互作用中对映选择性的表征方法研究进展

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分析科学学报 2016年第4期32卷 569-574页

作者：卢蓝蓝陈佳虹刘英菊肖治理张炽坚 sergei a.eremin 孙远明雷红涛华南农业大学食品学院广东省食品质量安全重点实验室广东广州510642 华南农业大学理学院生物材料研究所广东广州510642 A.N.Bach Institute of Biochemistry Russian Academy of SciencesMoscowRussia 119071

手性药物在临床上有着重要的影响,已成为人们关注的一个重点问题。本文对手性药物小分子与生物大分子间相互作用中对映选择性分析方法的研究进展作了概述,重点介绍了分子光谱分析法、等温滴定量热法、手性传感器、原子力显微镜、分子对... 详细信息

手性药物在临床上有着重要的影响,已成为人们关注的一个重点问题。本文对手性药物小分子与生物大分子间相互作用中对映选择性分析方法的研究进展作了概述,重点介绍了分子光谱分析法、等温滴定量热法、手性传感器、原子力显微镜、分子对接模拟等方法在手性药物小分子与生物大分子间对映选择性相互作用表征方法中的应用与进展。

关键词：手性药物生物大分子对映选择性