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百菌清人工抗原的制备与鉴定
引用本文:郭乃菲,高 兴,纪淑娟,于基成.百菌清人工抗原的制备与鉴定[J].食品科学,2010,31(13):214-218.
作者姓名:郭乃菲  高 兴  纪淑娟  于基成
作者单位:1.沈阳农业大学食品学院 2.大连民族学院生命科学学院
基金项目:辽宁省自然科学基金项目(20072916);大连民族学院人才引进项目(20086203)
摘    要:以杀菌剂百菌清为起始反应物,通过两步化学反应合成百菌清的衍生物(半抗原);采用活化酯法与混合酸酐法分别将半抗原与载体蛋白(牛血清蛋白、卵清蛋白)进行偶联,制备百菌清的人工抗原,同时通过免疫新西兰大白兔获得抗血清,用酶联免疫吸附法(ELISA)测定其效价。利用红外光谱(IR)、质谱(MS)、核磁共振波谱(NMR)对百菌清半抗原进行表征,并用紫外扫描的方法对人工抗原进行鉴定。结果表明,成功制备出百菌清半抗原及人工抗原,并且半抗原与载体蛋白的偶联比分别为27.86:1 和12.32:1,抗血清效价约为1:1.28 × 104。

关 键 词:百菌清  半抗原  人工抗原  鉴定  
收稿时间:2009-11-12

Preparation and Identification of Tetrachloroisophthalonethile Artificial Antigen
GUO Nai-fei,GAO Xing,JI Shu-juan,YU Ji-cheng.Preparation and Identification of Tetrachloroisophthalonethile Artificial Antigen[J].Food Science,2010,31(13):214-218.
Authors:GUO Nai-fei  GAO Xing  JI Shu-juan  YU Ji-cheng
Affiliation:1. College of Food Science, Shenyang Agricultural University, Shenyang 110161, China; 2. College of Life Science, Dalian Nationalities University, Dalian 116600, China
Abstract:In this work, tetrachloroisophthalonethile (TCPN) was derivatized into TCPN-COOH through two-step chemical reactions to prepare a hapten. TCPN artificial antigens, TCPN-COOH-BSA and TCPN-COOH-OVA were prepared from the hapten separately conjugated with carrier proteins, i.e., bovine serum albumin (BSA) and ovalbumin (OVA) by activated ester and mixed anhydride methods. An antisera was obtained through the immunization of New Zealand white rabbits using TCPNBSA. Antiserum titer was detected by enzyme-linked immunosorbent assay (ELISA). Meanwhile, TCPN-COOH and its intermediate product TCPN-OH were characterized by IR, MS and NMR methods, and TCPN-COOH-BSA and TCPNCOOH- OVA were qualitatively analyzed by UV/Vis spectrophotometry. TCPN-COOH, TCPN-COOH-BSA and TCPNCOOH- OVA were successfully synthesized, and the coupling ratios between TCPN-COOH and BSA or OVA were 27.86 and 12.32, respectively. The antiserum titer was about 1:1.28 × 104.
Keywords:tetrachloroisophthalonethile  hapten  artificial antigen  identification  
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