苄嘧磺隆人工抗原的合成与鉴定.pdf

※生物工程
品
2012.VoL33.No.19
苄嘧磺隆人工抗原的合成与鉴定
工磊,睢珂2,王志伟23,潘家荣*,赵杰,郑月明2
(1.中阳计量学院生命科学学院,浙.杭州310018:
2.中国农业科学院农产品加工研究所,北京100193;3.沈阳农业大学食品学院,辽予宇沈阻110866
摘要:以2-甲酸甲恉苄磺嘞胺和現珀酸为原料合成半抗原( hapten),经薄层层析和H-NMR際定后,采用混合酸
分别与作血清蛋白(3SA)和卵清白蛋白(OVA)偶联获得免疫原( halen-IBSA)和包被原( hapten-OVA),紫外光谱分析法计
算得其偶咲比分别为8:1和13:1,初步说明具有较好免疫原性。通过动物免疫实验获得了效价达1:32000的阳性血清,采
用间接悔联免疫吸附檢測( ELSA)法檢測抗体的ICs和C。值为05032mg和004mgL,表明人工抗原合成成功。
关键词:嘧磺隆:磺隆除草剂;联兔疫败附檢
Synthcsis and Idcntification of Artificial Antigcn for Bcnsulfuron
WANG Lei, SU Ke, WANG Zhi-wet", PAN Jia-rong", ZLAO", ZENG Yue-mmg
(1. College of Iife Sciences, China iliang [niversity. Hangzhou 310018, China;
2. Agro-food Processing Institute, Chinese Academy of Agricultural Sciences, Beijing 100193, China
3. College of Food Science, Shenyang Agricultural University, Shenyang 110866, China
Abstract: In this study, a apen was s thesize from methyl 2 -(sulfamoyl)methyl benzoate and succinic anhydride. The
synthetic hapten was idcntified by thin layer chromatography and H-NMR spectroscopy. An immune antigen(hapten-bsa
and a coating antigcn(haptcn-va)were prepared through scparatc conjugation of the hapten with 1SA and (VA by mixcd
anhydride method. The conjugation ratios of the Immune antigen and coating antigen were calculated to be 8: 1 and 13: 1
based on UV spectroscopic analysis, respectively, suggesting their good immunogenicity. Positive sera with a titer of 1: 32000
were oblained by animal immunization trials. I'he IC o and Ico o[ antibody were determined by indireet FIIS assay to be
0. 5032 mg/L and 0.0024 mg L, respectivcly. Therefore, the synthesis of artificial antigen is successul
Key words: bensulfuron; sulfonylurea herbicide; ELISA
中图分类号:Q914.5
献标识码:A
文章编号:1002-630(2012)19-0223-04
横隆类除草剂是口前世界上最大一类除草剂,自从期延长的特点,造成对当地农作物及环境的危害和损失
杜邦公可1982年首次开发出麦出除草剂氯磺隆以后,通事件频繁报道,公众环保和健康意识的提高也迫使我们
过在磺戲脲灲端及桥上进行修饰等衍生化方法,目前要加强对其残留的监测研究。欧盟已经対饮用水和果蔬
已有近40个品种商品化。横降类除草剂通过抑制乙酰
巾做出最低限量标准为0.05mgkg?
酸合成酶( acetolactate synthase,AIS)、乙酸羟酸合成
传统的隆类除草剂残留检测方法有气相色谱
( acetohydroxyacid synthase,AHAS),从而破坏蛋白质合液相色谱和毛細管也泳等方法。气相色法由于
成、干扰DNA合成和细胞分裂与生长,使杂草正常生长脉类发性低和热不稳定需要衍生化;液相色谱配合质
受到破坏而死心山。它因高效、用量少、声性低及半衰谱及固相微萃取等方法虽能对样品中横酰脲类除草剂进
时间短而广泛应げ于水稱、小麦、玉米等大出作物及花行灵敏准确检测,国内外已有大量文献研究报道1的
生、马铃薯瓜果蔬菜以。但是磺隆类化合物因不易挥但试剂有毒且分析成本高昂; Menne、 Garcialb等用毛
发、不易光解、有机吸附系数低、随上壤pH值上升半衰细管电泳方法対上壤样品巾磺隆类除草剂进行分析岍究
收稿口期:2011-09-26
基金项目:“十?”国家科技支撑计划项目(200D3AD9303-Z04)
信行:家一男,究贝,し、研究方向为检!技不、
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