呋喃唑酮残留标示物人工抗原合成与抗体制备

食品科学 ›› 2008, Vol. 29 ›› Issue (12): 444-448.

呋喃唑酮残留标示物人工抗原合成与抗体制备

常超, 伍金娥, 袁宗辉

武汉工业学院食品科学与工程学院；华中农业大学国家兽药残留基准实验室,农业部食品安全评价重点开放实验室；

出版日期:2008-12-15 发布日期:2011-12-08

Antigens Synthesis and Antibodies Preparation of Furazolidone Marker Residue

CHANG Chao, WU Jin-E, YUAN Zong-Hui

1.College of Food Science and Engineering,Wuhan Polytechnic University,Wuhan 430023,China;2.National Reference Laboratory of Veterinary Drug Residues,Key Laboratory of Food Safety Evaluation,Ministry of Agriculture,Huazhong Agricultural University,Wuhan 430070,China

Online:2008-12-15 Published:2011-12-08

摘要/Abstract

摘要： 以乙醇肼和碳酸二甲酯为原料,合成2种半抗原3-氨基-2-恶唑烷酮(AOZ)和3-(4-羧基苯亚甲基)-氨基-2-恶唑烷酮(CPAOZ),经戊二醛法和碳二亚胺法将半抗原与牛血清蛋白(BSA)偶联合成2种人工抗原AOZ-BSA和CPAOZ-BSA,产物经紫外光谱扫描分析鉴定。用2种人工抗原采取不同程序免疫新西兰大白兔,获得8组多克隆抗体。AOZ-BSA抗体效价1:0.8×104～1:3.2×104,对AOZ灵敏度低,IC50均大于100μg/ml。CPAOZ-BSA抗体效价1:1.6×105～1:6.4×105,对AOZ与苯甲醛衍生物N-苯亚甲基-3-氨基-2-恶唑烷酮(PAOZ)灵敏度高,IC500.91～11.56ng/ml,特异性好,仅与呋喃唑酮(交叉反应率6.15%)、3-(2-硝基苯亚甲基)-氨基-2-恶唑烷酮(NPAOZ,交叉反应率5.73%)有交叉反应,与衍生试剂苯甲醛和其他药物没有交叉反应(交叉反应率<0.01%)。本研究制备出AOZ特异性抗体,为我国开发AOZ酶联免疫试剂盒奠定基础。

关键词: 呋喃唑酮, 半抗原, 人工抗原, 抗体

Abstract: Two haptens,3-amino-2-oxazolidinone(AOZ) and 3-{[(4-carboxyphenyl) methylene]amino}-2-oxazolidinone(CPAOZ),were prepared by using 2-hydroxyethyl hydrazine and diethyl carbonate as raw materials. The haptens were conjugated to bovine serum albumin(BSA) by the glutaraldehyde and the carbodiimide procedures respectively. The conjugates AOZ-BSA and CPAOZ-BSA were identified with ultraviolet spectrum. Eight polyclonal antibodies were produced by the various immunization of New Zealand rabbits with the conjugates. The antibodies produced with AOZ-BSA conjugate show the titers in the range of 1:0.8×104 to 1:3.2×104 and the IC50 values of more than 100 μg/ml,respectively. These antibodies are not specific for the AOZ moiety. The antibodies produced with CPAOZ-BSA conjugate show the titers in range of 1:1.6×105 to 1:6.4×105,the IC50 values in range of 0.91 to 11.56 ng/ml,and cross-reactivity of 6.15% with furazolidone and 5.17% with 3-{[(2-nitrophenyl)methylene]amino}-2-oxazolidinone(NPAOZ),respectively. These antibodies are highly specific for a benzaldehyde derivative of AOZ,N-(phenzlidene)-3-amino-2-oxazolidinone(PAOZ) and do not cross-react with various veterinary drugs.

Key words: furazolidone, hapten, antigen, antibody

常超, 伍金娥, 袁宗辉. 呋喃唑酮残留标示物人工抗原合成与抗体制备[J]. 食品科学, 2008, 29(12): 444-448.

CHANG Chao, WU Jin-E, YUAN Zong-Hui. Antigens Synthesis and Antibodies Preparation of Furazolidone Marker Residue[J]. FOOD SCIENCE, 2008, 29(12): 444-448.

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