邓安平
小分子化合物的免疫分析化学方法学及其应用的研究
个性化签名
- 姓名:邓安平
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学术头衔:
博士生导师
- 职称:-
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学科领域:
分析化学
- 研究兴趣:小分子化合物的免疫分析化学方法学及其应用的研究
邓安平,教授、博士生导师,1984 南京大学化学系 学士; 1990 华西医科大学生物化学专业 硕士; 1999 捷克 Masaryk 大学分析化学专业 博士。 1984-1987 华西 医科大学 化学教研室 助教; 1990-1995 华西 医科大学 卫生检测教研室 讲师; 2000-2002 台湾中山大学化学系 博士后; 2002-2004 德国慕尼黑工业大学化学系 博士后; 2004.7 至今,四川大学化学学院 教授 博士生导师。 曾 到西班牙奥维耶多大学、韩国庆尚南道大学、德国明斯特大学 、 香港科技大学 、美国肯塔基大学进行 1-3 个月的学术交流。
科研方向主要为小分子化合物的免疫分析化学方法学及其应用的研究 , 包括以下几个方面:(1) 免疫分析方法 学 的研究,如 酶联免疫吸附分析 、 化学发光免疫分析 、 感耦合等离子体 - 质谱 - 免疫吸附分析 、 激光诱导荧光免疫分析 、 免疫传感器 、 流动注射免疫分析等。(2) 免疫分析方法应用的研究 , 测定环境、食品、药物、生物等样品中的小分子物质, 如农药、工业污染物、环境激素、违禁食品添加剂、毒素、药物、多肽等。 (3) 对于成熟的免疫分析方法,研制、开发成免疫分析试剂药盒,并使之商品化。(4) 功能生物材料的制备、表征及应用的研究,如磁性纳米材科、分子印迹聚合物。(5) 高效 分离方法的研究,如 免疫亲合层析分离技术 、分子印迹技术 用于痕量小分子化合物和中药有效成份的分离、富集。
在国内外学术期刊上发表科研论文四十多篇,其中两篇论文( Anal. Chem. 2002 , 74 , 2617-2621 : Environ. Sci. Technol. 2003 , 37 , 3422-3429) 单篇他引均超过 16 次。 参与研发的多氯联苯、磺胺二甲基嘧啶、 2 , 4- 二氯苯氧基乙酸、阿特拉津、西玛津等免疫试剂盒,已先后在欧洲上市。申请发明专利一项。在研科研项目:1、国家自然科学基金 No. 20675054;2、教育部留学回国人员科研启动基金 No. 200 6331-11-3;3、四川大学振兴计划。
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成果阅读
248
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成果数
7
邓安平, Diana Matschulat, Anping Deng, † Reinhard Niessner and Dietmar Knopp*
,-0001,():
-1年11月30日
In Europe, a limit value of 10ng L21 was set by the European Commission for benzo[a]pyrene(B[a]P) in water intended for human consumption (Council Directive 98/83/EC) and, therefore,sensitive and reliable methods are needed to evaluate its presence. We report here on thedevelopment of a highly sensitive indirect competitive ELISA for the detection of B[a]P in potablewater. Fourteen monoclonal antibodies were generated in mice using novel B[a]P derivatives. Theimmunoassay with the least interference and the best sensitivity was optimized and characterized.As co-solvent, ten percent methanol (v/v) was determined as the optimum concentration for B[a]Psolubilization for use with the developed ELISA. With the purified antibody (clone 22F12) theaverage IC50 for B[a]P and corresponding detection limit at a signal:noise (S/N) ratio of 3 was65 ng L21 and 24 ng L21, respectively. From the 16 EPA-designated PAHs, only chrysene,indeno[1,2,3-cd]pyrene, and benzo[b]fluoranthene showed a cross-reactivity (CR) higher than20%. No CR was observed for two- and three-ringed aromatics as well as dibenz[ah]anthraceneand benzo[ghi]perylene. The effect of pH value (range 6.5-9.5), ionic strength (specific electricconductivity 1 mS cm21–2.5mS cm21), and inorganic ions (sodium, copper, iron, aluminium,manganese, chloride, sulfate, nitrate, and nitrite at maximum permissible levels according to theCouncil Directive) on both signal and sensitivity of the ELISA was studied. No significantinfluence of these parameters on the ELISA competition curve was found. We suggest that theoptimized ELISA can be used to monitor potable water samples without previous extraction fromthe samples. The assay should facilitate the cleanup of B[a]P contaminated sites where B[a]P levelsfall close to the limit value of the new drinking water directive.
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【期刊论文】Residue Analysis of thePharmaceutical Diclofenac inDifferent Water Types Using ELISAand GC-MS
邓安平, ANPING DENG, #MARKUS HIMMELSBACH, §QING-ZHIZHU, † SIEGFRIED FREY, |MANF RED SENGL, |WOLFGANG BUCHBERGER, §REINHARD NIESSNER, ‡ AND DIETMAR KNOPP*, ‡
Environ. Sci. Technol. 2003, 37, 3422-3429,-0001,():
-1年11月30日
A highly sensitive and specific indirect competitiveenzyme-linked immunosorbent assay (ELISA) for thedetermination of diclofenac in water samples was developed.With pure water, the limit of detection (LOD, S/N ) 3)and IC50 were found to be 6 ng/L and 60 ng/L, respectively.The analytical working range was about 20-400 ng/L.Highest cross-reactivity (CR) of 26 tested pharmaceuticals,metabolites, and pesticides was found for 5-hydroxydiclofenac(100%). Other estimated values were well below4% and, therefore, are negligible. The assay was appliedfor the determination of diclofenac in tap and surface watersamples as well as wastewater collected at 20 sewagetreatment plants (STPs) in Austria and Germany. Humicsubstances were identified as main interference in surfacewater. Wastewater samples which were only submittedto filtration and dilution yielded about 25% higher diclofenacconcentrations using the ELISA compared to GC-MS.However, the ELISA turned out to be a simple, inexpensive,and accurate method for the determination of diclofenacboth in influent and effluent wastewater after rather simplesample preparation, i.e., filtration, acidification, andreadjustment to neutral pH-value, and at least 10-folddilution with pure water.
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邓安平, An-Ping Denga, , Jiin-Tsuey Chengb, Hsuan-Jung Huanga, ∗
Analytica Chimica Acta 461(2002)49-55,-0001,():
-1年11月30日
An amperometric immunoassay was developed by coupling the enzyme-linked immunosorbent assay (ELISA) microtiterplatesystem with a polyaniline-perfluorosulfonated ionomer composite (PA/NF) electrode incorporated flowinjection analysis(FIA) system and used for the analysis of Tal 1 protein, found in leukemic T cell. Rabbit polyclonal antibody (pAb) against Tal1 and urease–pAb were used, respectively as the captured protein and enzyme labeled conjugate for sandwich immunoassay ofTal 1. Characteristics of the PA/NF electrode such as reproducibility, stability and sensitivity were studied. The detection limitsof the PA/NF electrode for NH4+ and urease were found to be 5_M and 0.05 nM, respectively. Assay conditions such as theamount of pAb needed for coating the plate, the concentration of urease–pAb conjugate appropriate for the immunoreactionand the incubation time for urea to react with the bound urease–pAb in the microtiter-wells were also studied. A detectionlimit as lower as 0.5 ng/ml and a dynamic range of 1.0-100 ng/ml were found for the immunoassay of Tal 1 protein with thedeveloped immunoassay system.
Urease, Amperometric immunoassay, FIA, Polyaniline, Nafion, Tal 1 protein
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邓安平, An-Ping Denga, Hui-Tao Liu b, Shiuh-Jen Jiang a, ∗, Hsuan-Jung Huanga, , Chi-Wi Onga
Analytica Chimica Acta 472(2002)55-61,-0001,():
-1年11月30日
The high sensitive dynamic reaction cell inductively coupled plasma mass spectrometry (DRC ICP-MS)-based immunoassayusing ferrocene (Fc) tethered hydroxysuccinimide ester as label for the determination of 2,4-dichlorophenoxyacetic acid(2,4-D) was presented. Ferrocene tethered hydroxysuccinimide ester was directly or via horse radish peroxidase (HRP) as abridge coupled to monoclonal antibody (mAb) of 2,4-D. Competitive immunoreactions were completed in microtiter plateand the signal of 56Fe in the bound ferrocene labeled conjugate was detected by DRC ICP-MS. The potentially interfering40Ar16O+ at the iron mass m/z 56 was reduced in intensity significantly by using NH3 as the reaction gas. The optimizationprocess of the immunoassay was greatly simplified in the aid of enzyme linked immunosorbent assay (ELISA) procedure. The2,4-D was determined in the dynamic range of 0.1–1000 ng/ml and the detection limits of the assays using the two ferrocenelabeled conjugates were found to be 0.044 and 0.055 ng/ml, respectively. The relative standard deviation for six measurementswere within 5.5–15.3%.
Immunoassay, Inductively coupled plasma mass spectrometry, Ferrocene labeled conjugate, 2,, 4-Dichlorophenoxyacetic acid
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【期刊论文】Immunoassay with a Microtiter Plate Incorporated Multichannel Electrochemical Detection System
邓安平, Tzyh-Chyang Tang, † Anping Deng, ‡, § and Hsuan-Jung Huang*
Anal. Chem. 2002, 74, 2617-2621,-0001,():
-1年11月30日
To extend the application of a multichannel electrochemical detector (MED) for immunoassay, a MED system consisting of 8 sets of Pt electrodes in an arrangement fitted with the dimensions of a row of microtiter wells in a microtiter plate and a microcomputer-assisted 16-channel potentiostat was constructed. With this developed MED system, electroactive enzymatic products produced in eight microtiter wells can be analyzed simultaneously with a developed amperometric procedure. To demonstrate the applicability of the MED system for immunoassay, an immunosystem containing rabbit-IgG and alkaline phosphatase-conjugated goat anti rabbit-IgG was studied. From the dynamic range of 10-1000 ng/mL (0.064-6.4pM) and the detection limit of 1.0 ng/mL (6.4pM) obtained, the developed MED shows a tremendous improvement in sensitivity, detection limit, and efficiency compared with that obtained from conventional electrochemical immunoassay.
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【期刊论文】Direct competitive immunoassays for the coplanar polychlorinated biphenyls
邓安平, Milan Fránek a, ∗, Anping Denga, Vladimir Kolar a, Jaromir Socha b
Analytica Chimica Acta 444(2001)131-142,-0001,():
-1年11月30日
lic acid spacers at para-position were derived from coplanar 3,3,4,4-tetrachlorobiphenyl and 3,3_,4,4_,5-pentachlorobiphenyl (IUPAC No. 77 (PCB 77) and 126 (PCB 126), respectively). It appeared from the results that the length of the spacer did not affect significantly the sensitivity of the assays established. The direct ELISAs using the antibodies in combination with various hapten–peroxidase tracers showed IC50-values for polychlorinated biphenyl (PCB) 77 and 126 of 1.3–19.2μg l−1. The ELISAs were tested for their cross-reactivity profiles using 20 selected congeners. The assays were highly specific for non-ortho-substituted (coplanar) congeners and did not recognize the more abundant but less toxic non-coplanar PCBcongeners or organochlorine compounds. The superior assaywas found for the antibody againstPCB126-(CH2)3-hapten mimic–bovine serum albumin (BSA) in combination with PCB 77-CH2-hapten mimic-peroxidase tracer. The IC50-values for the assay to PCB 77 and 126 were 2.0 and 5.2μg l−1, respectively; the highest cross-reactivity values (PCB 126=100%) achieved for non-coplanar structures and metabolites were only 1.3 and 1.0% for 2,3,3,4,4,5,5 heptachlorobiphenyl (PCB 189) and PCB 77 ether, respectively. In order to interpret measured data in terms of analytical or toxicological equivalents, a reliable relationship between assay responses and the coplanar congener content of Aroclor-contaminated samples should be verified and validated.
Immunoassays, Immunogen, Tracer, Spacer, Coplanar PCBs, Cross-reactivity, Dose-response
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邓安平, Milan Frane*, Anping Deng, Vladimir Kolar
Analytica Chimica Acta 412(2000)19-27,-0001,():
-1年11月30日
A sequential injection instrument (ALITEA, USA) with a photometric and fluorometric detection unit S2000 (Ocean Optics) was employed for the development of flow injection immunoanalysis (FIIA). The monoclonal antibodies against atrazine, simazine and 2,4-D were immobilized on aminopropyl glass particles by means of avidin/biotin system and packed in plexiglass column of 18ml volume. Assay characteristics for individual antibody-reactors and regeneration effectivities for acid and alkaline solutions are described. An attempt to prepare a functional mixed antibody-reactor has not achieved success since regeneration conditions found for individual reactors were not compatible with one performance protocol.
Flow injection immunoassay, Monoclonal antibodies, Immunoreactor, Regeneration, Calibration
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