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Preparation of monoclonal antibody based indirect competitive ELISA for detecting 19-nortestosterone residue

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Chinese Science Bulletin 2011年第25期56卷 2698-2705页

作者： jiang jinqing ZHANG HaiTang FAN GuoYing MA JinYou WANG ZiLiang WANG JianHua College of Animal Sciences Henan Institute of Science and Technology Xinxiang 453003 China College of Veterinary Medicine Northwest A&F University Yangling 712100 China

19-Nortestosterone (NT) has been illegally used in horse racing to boost physical performance, and in animal husbandry to accelerate weight gain. To monitor the abuse of NT, our goal was to develop a commercial enzyme linked immunosorbent assay (ELISA) kit. For this purpose, hybridomas were prepared by fusing NS0 mouse myeloma cells with splenocytes isolated from immunized BALB/c mouse. Noncompetitive and competitive indirect ELISA were used to screen positive cell clones. To optimize the indirect competitive ELISA (icELISA) method, various methanol concentrations in assay buffer were evaluated. Matrix effects in urine and spiking test were also investigated. Finally, five hybridoma cell lines named NT-1, NT-2, NT-3, NT-4 and NT-5 were screened out. The corresponding monoclonal antibodies (mAbs) were of the IgG 1 isotype with a k light chain, and the antibody affinity of all mAbs were between 2.6×10 9 and 4.7×10 9 L/mol. The titer and IC 50 values of purified ascites were in the range of 0.64×10 5 2.56×10 5 and 0.55-1.0 ng/mL, respectively. Based on the NT-1 hybridoma, a heterologous icELISA method was developed for the quantitative detection of NT in cattle urine. The dynamic range was from 0.004 to 85.8 ng/mL, with a detection limit for the assay and IC 50 values of 0.002 and 0.55 ng/mL, respectively. Except for a high cross-reactivity (62%) to α-NT, negligible cross-reactivity to other compounds was observed. After optimization, 10% of methanol was used in the assay buffer, and a 20-fold dilution in cattle urine gave an inhibition curve almost the same as that in phosphate buffered saline. The correlation coefficient between the established icELISA and LC-MS/MS method was 0.9871. The results showed that the established heterologous icELISA method provides an excellent alternative for the detection of NT residues in food producing animals.

关键词： ELISA检测间接ELISA 单克隆抗体竞争力酶联免疫吸附试验残留睾酮 ELISA方法

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结构稳定性的线性与非线性分析

西北工业大学学报 1987年第3期5卷 335-344页

作者：竺润祥姜晋庆西北工业大学西北工业大学副教授

本文讨论采用有限元法对结构进行稳定性分析方法,侧重于介绍非线性分析方法。采用伴有各种牛顿型和线性搜索的组合弧长法可以顺利描述具有极值点的非线性基本平衡路径。在求解平衡点同时,附加计算该点的行列式值、特征值或刚度特性参数... 详细信息

本文讨论采用有限元法对结构进行稳定性分析方法,侧重于介绍非线性分析方法。采用伴有各种牛顿型和线性搜索的组合弧长法可以顺利描述具有极值点的非线性基本平衡路径。在求解平衡点同时,附加计算该点的行列式值、特征值或刚度特性参数,根据稳定准则,通过插值可确定临界点。同时,本文还介绍了超屈曲路径描述方法.由计算实例表明,本文提供的稳定性分析方法简单而有效,可供工程设计参考。

关键词：平衡路径特征方程结构稳定性分叉点极值点行列式值非线性分析数学分析

Effects of Germ Line Transmission in Cloned Calves Derived from In Vitro-Transfected Somatic Cells

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Effects of Germ Line Transmission in Cloned Calves Derived f...

2011 International Conference on Engineering and Information Management (ICEIM 2011)

作者： An Zhixing jiang jinqing Zhang Xiaojian Wang Qinghua Zhang Jinzhou College of Animal Science Henan Institute of Science and Technology

In vitro transfection of cultured cells combined with nuclear transfer currently is the most effective procedure to produce transgenic livestock. In the present study, bovine primary fetal fibroblasts were transfected with a green fluorescent protein (GFP) reporter transgene and used as nuclear donor cells in oocyte reconstructions. To examine the role of host cytoplasm on transgene expression and developmental outcome, GFP-expressing fibroblasts were fused to oocytes reconstructed either metaphase or telophase activation, and PCR technology was also employed. The results showed that GFP became detectable at the 8-to 16-cell stage, approximately 80 h after reconstruction, and remained positive at all later stages. Embryonic development to the blastocyst stage was not significantly different among metaphase and telophase groups. Therefore, GFP transgene technology can be used to select embryoes derived from transgenic animals.

关键词： Transgenic animal Cloned calves Embryo In vitro fertilization Green fluorescent protein

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固体火箭发动机连接结构接触应力研究

推进技术 1997年第4期18卷 54-57页

作者：南宫自军姜晋庆张铎西北工业大学航天工程学院西安710072

应用接触理论和有限元方法，分析固体火箭发动机法兰连接结构的接触问题。建立了法兰连接结构的有限元模型，充分考虑了连接螺栓预紧力对结构刚度的影响，将其合理地转化为等效节点初载荷。推导出在法兰对接面上有气密性要求时求解最佳... 详细信息

应用接触理论和有限元方法，分析固体火箭发动机法兰连接结构的接触问题。建立了法兰连接结构的有限元模型，充分考虑了连接螺栓预紧力对结构刚度的影响，将其合理地转化为等效节点初载荷。推导出在法兰对接面上有气密性要求时求解最佳预紧力的公式。最后通过实例进行了接触应力计算。结果表明，采用的分析方法和由此得出的结论具有一定的工程意义。

关键词：封头螺栓连接接触应力固体火箭发动机

矿井主要通风机反风集中远程控制系统研究与应用

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煤矿现代化 2019年第1期28卷 76-77页

作者：蒋金庆山东东山王楼煤矿有限公司山东济宁272063

为了实现主要通风机反风集中远程控制,分析了反风操作流程及影响因素,分析表明防爆井盖未实现远程控制和各远程控制系统单一操作是主要影响因素。从防爆井盖远程控制和对各控制系统对接集成入手,形成了主要通风机反风集中远程控制系统,... 详细信息

为了实现主要通风机反风集中远程控制,分析了反风操作流程及影响因素,分析表明防爆井盖未实现远程控制和各远程控制系统单一操作是主要影响因素。从防爆井盖远程控制和对各控制系统对接集成入手,形成了主要通风机反风集中远程控制系统,实现了无人值守集中远程控制反风。

关键词：主要通风机反风集中远程控制

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室内空气污染及其控制对策

绍兴文理学院学报（自然科学版） 2005年第4期25卷 42-45页

作者：蒋锦青申秀英东阳市环保局浙江东阳322100 浙江师范大学化学与生命科学院浙江金华321004

从室内空气污染物主要来源、污染物及其危害入手，针对我国国情和以往我国室内空气质量控制存在的问题，从社会、技术等角度提出行政管理对策和技术防治对策．

关键词：室内空气污染防治对策

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猪新发冠状病毒研究进展

畜牧兽医学报 2020年第10期51卷 2359-2366页

作者：李任峰卢晓辉姜金庆王自良河南科技学院动物科技学院河南省高校动物疫病防控科技创新团队新乡453003 新乡市动物疫病预防控制中心新乡453003

当前,由新型冠状病毒(SARS-CoV-2)导致的新型冠状病毒肺炎(COVID-19)疫情正在全球暴发大流行,这也引发业界对于动物源性冠状病毒的高度关注。猪德尔塔冠状病毒(PDCoV)和猪急性腹泻综合征冠状病毒(SADS-CoV)是近年来新发现的猪冠状病毒,... 详细信息

当前,由新型冠状病毒(SARS-CoV-2)导致的新型冠状病毒肺炎(COVID-19)疫情正在全球暴发大流行,这也引发业界对于动物源性冠状病毒的高度关注。猪德尔塔冠状病毒(PDCoV)和猪急性腹泻综合征冠状病毒(SADS-CoV)是近年来新发现的猪冠状病毒,不但严重危害到养猪业,对公共卫生安全也具有潜在威胁风险。本文结合国内外现有文献,从PDCoV和SADS-CoV的病原学、病毒起源与进化、致病性以及检测诊断技术等方面进行综述,并提出展望,以拓展对SARS-CoV-2的认识,并为PDCoV和SADS-CoV的后续研究提供参考借鉴。

关键词：猪新发冠状病毒猪德尔塔冠状病毒猪急性腹泻综合征冠状病毒