|
|
|
题名
|
作者
|
年代
|
出处
|
被引量
|
| 1 | 雌二醇间接竞争酶联免疫吸附方法的建立显示文摘建立食品中雌二醇免疫分析方法。将雌二醇进行结构改造合成半抗原并经过核磁氢谱和红外光谱验证,再采用活泼酯法与载体BSA和OVA偶联制备人工抗原,免疫兔子制备多克隆抗体,经过反应条件优化建立间接竞争酶联免疫吸附检测方法。结果表明:IC50为3.9ng/mL,检测限为0.3ng/mL,交叉反应率均小于1.21%。检测方法具有高灵敏度和特异性,可应用于食品中雌二醇检测试剂盒的研制。 | 刘珒 雷红涛 孙远明 李振峰 杨金易 沈玉栋 王弘 曾道平 | 2012 | 食品科学2012，33，8： | 7 |
| 2 | 19-去甲睾酮单克隆抗体的筛选及icELISA方法的建立显示文摘采用细胞融合技术筛选抗19-去甲睾酮(nortestosterone,NT)单克隆抗体(monoclonal antibody,mAb)杂交瘤细胞株,体内诱性腹水法制备NT mAb,建立异源性间接竞争ELISA(indirect competitive ELISA,icELISA)标准曲线,并对其特异性、理化因素的影响和基质效应性进行探讨.结果表明,筛选的5株杂交瘤细胞分泌的单抗腹水效价为0.64×105~2.56×105,半数抑制浓度IC50为0.55~1.0ng/mL,抗体为IgG1亚型,具k轻链,亲和常数Ka为2.6×109~4.7×109L/mol.建立的icELISA标准曲线在磷酸盐缓冲液(phosphate buffered saline,PBS)中的线性检测范围为0.004~85.8ng/mL,IC50值为0.55ng/mL,检测限(limit of detection,LOD)为0.002ng/mL;抗体与17a-去甲睾酮的交叉反应率(cross-reactivity,CR)为62%,与其他化合物无CR.标准品稀释液中甲醇的最佳含量为10%,牛尿1:20倍稀释可消除基质干扰,ELISA方法和液质联用检测(LC-MS/MS)的相关性良好(R2=0.9871). | 姜金庆 张海棠 范国英 马金友 王自良 王建华 | 2011 | 科学通报2011，56，20： | 5 |
| 3 | Preparation of monoclonal antibody based indirect competitive ELISA for detecting 19-nortestosterone residue显示文摘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. | JIANG JinQing ZHANG HaiTang FAN GuoYing MA JinYou WANG ZiLiang WANG JianHua | 2011 | Chinese Science Bulletin2011，56，25： | 4 |
| 4 | 诺氟沙星单克隆抗体的制备及初步应用显示文摘采用优化的碳二亚胺法制备诺氟沙星(NOR)人工抗原,紫外扫描和红外光谱图表明人工抗原偶联成功。采用细胞融合技术筛选抗NOR单克隆抗体(NORmAb)杂交瘤细胞株,体内诱生腹水法制备NORmAb,建立间接竞争ELISA(icELISA)标准曲线。结果表明,筛选的3株杂交瘤细胞分泌的抗体为IgG1亚型,具k轻链;建立的icELISA标准曲线在PBS中的的线性检测范围为0.04~43.6ng/mL,IC50值为0.62ng/mL,检测限(LOD)为0.02ng/mL;抗体与环丙沙星(87.2%)、培氟沙星(76.9%)、依诺沙星(66.8%)、恩诺沙星(48.6%)和洛美沙星(36.6%)有较高的交叉反应率;禽肉基质中NOR添加回收率满足检测要求。 | 李超英 姜金庆 | 2011 | 食品工业科技2011，32，9： | 3 |
| 5 | Production and Characterisation of Monoclonal Antibodies against 19-Nortestosterone显示文摘Objective To produce anti‐19‐Nortestosterone (NT) monoclonal antibodies and identify their immunological characteristics. Methods Hybridomas were prepared by fusing NS0 mouse myeloma cells with splenocytes isolated from immunized BALB/c mice. Noncompetitive and competitive indirect ELISA were employed to screen positive cell clones. A caprylic acid ammonium sulphate (CAAP) method was used to purify NT mAb, and the Batty saturation method was used to determine the affinity constant (Kaff). Results Five hybridoma cell lines, named NT‐1, NT‐2, NT‐3, NT‐4, and NT‐5, were identified and their corresponding mAbs were of the IgG 1 isotype with a k light chain. The Kaffs of all mAbs were between 2.6 and 4.7×10 9 L/mol. The titers and IC 50 values of purified ascite fluids were in the range of (0.64-2.56)×10 5 and (0.55-1.0) ng/mL, respectively. Of all the cross‐reacting steroids, α‐NT was the most reactive with the mAbs at 62% with NT‐1 mAb and 64% with NT‐2 mAb. Negligible cross‐reactivity (<0.01%) with other steroids was observed. Conclusion The establishment of these hybridomas allows the potential development of a rapid test kit, and may provide an alternative method for the detection of NT residues in food producing animals. | JIANQ JinQing ZHANG HaiTang YAO SiXin WANG ZiLiang WANG JianHua | 2011 | Biomedical and Environmental Sciences2011，24，2： | 2 |
| 6 | 抗诺氟沙星单克隆抗体的制备及其性质的研究显示文摘为了监测诺氟沙星抗生素的滥用,我们开发了一种基于单克隆抗体的酶联免疫方法。将小分子抗原诺氟沙星NOR通过碳二亚胺法与载体蛋白BSA、OVA进行偶联。得到完全抗原NOR-BSA和NOR-OVA。用NOR-BSA对3只雌性小鼠进行免疫后,利用细胞融合技术制备杂交瘤细胞,用间接ELISA法和间接竞争ELISA法筛选阳性细胞进行克隆,得到3株稳定分泌抗体的单克隆细胞株,分别命名为7C4,8G3,11G7。检测细胞的上清液和纯化后的腹水效价,分别为1:512,1:1024,1:1024和1:128000,1:128000,1:128000。对三株细胞的亚型鉴定为IgG2b,IgG2b,IgG1。建立的标准竞争曲线计算得IC50值为31.225 ng,其检测线性范围为5.011-630.95 ng/mL,最低检出量为2.75 ng。亲和力常数Kaff为4.6×10~8。诺氟沙星单克隆抗体与环丙沙星(27.22%)、恩诺沙星(11.29%)、奥比沙星(11.03%)、氧氟沙星(9.73%)、达氟沙星(7.38%)、沙拉沙星(2.57%)、二氟沙星(0.61%)几种抗菌药有较低的交叉反应率。 | 徐田丽 苏国成 江晓颖 孙雪珂 江峰 | 2017 | 科技风2017，，12： | 0 |
您所在的机构暂未开通此部分的功能，请联系管理员