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苯巴比妥人工避免疫原分松及抗体特点切磋

类别:皇冠比分网 来源:皇冠比分网|皇冠比分|皇冠比分官网| welcome!! 发布人:locoy 发布时间:2019-02-21 06:23 热度:°
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  干者:不知

  摘要:将苯巴比妥(PB)经硝募化和恢骈两步募化学修饰后,在其苯环上伸入活性基团弄氨基,分松具拥有半抗原构造特点的对氨基苯巴比妥(pAPB);用重氮募化法将pAPB偶联于载体蛋清BSA和OVA上,分松人工避免疫原BSA-pAPB和包被抗原OVA-pAPB,用红外面(IR)、紫外面(UV)、SDS-PAGE终止评判,铰算分儿子结合比;用BSA-pAPB避免疫新正西兰白兔和SPF雏鸡,直接ELISA测定多克隆抗体(pAb)效价,阻断ELISA评判其敏理性,WestGold层析试验和提交叉反应考验评判其特异性。结实标注皓,BSA-pAPB偶联成,分儿子结合比为1∶19;得到了低价、敏感、特异的pAb,为PB残剩避免疫学检测方法的确立奠定了基础。

  皇冠比分:苯巴比妥;半抗原;人工避免疫原;抗体;评判

  中图分类号:S859.79+1;S852.4+3;TS201.6 文件标注识码:A 文字编号:0439-8114(2008)07-0815-04

  Study on Synthesis of Artificial Immunogen and Characteristics of Polyclonal Antibody of Phenobarbital

  ZHANG Hai-tang,WANG Zi-liang,WANG Yan-rong,ZHONG Hua,FAN Guo-ying

  (College of Animal Science, Henan Institute of Science and Technology, Xinxiang 453003,Henan, China)

  Abstract: The active group amido was introduced into phenyl of PB and formed p-amidophenobarbital(pAPB) which had hapten structure by chemical modification of nitration and deoxidization. Diazotization was used to conjugate pAPB to carrier protein BSA and obtained artificial immunogen BSA-pAPB, the coating antigen OVA-pAPB was obtained in the same way. IR, UV and SDS-PAGE were used to identify PB artificial immunogen. New Zealand white rabbits and SPF chickens were immunized with BSA-pAPB, the titre of polyclonal antibody(pAb) was measured by indirect ELISA, the sensitivity of the pAb was identified by blocking ELISA and the specificity of pAb was identified by experiments of WestGold immuno chromatography and cross-reaction. The results showed that PB artificial immunogen was synthesized successfully and its conjugation ratio of pAPB to BSA was about 19∶1, the high-titer, sensitive and specific PB pAb were produced in sera of Balb/C mice immunized with BSA-pAPB, which made it possible to establish immunoassay of PB residues in animal food.

  Key words: phenobarbital; hapten; artificial immunogen; antibody; identification

  苯巴比妥(Phenobarbital,PB)佩名鲁米那(Luminal)、迦地那(Gardenal),首要成分为5-乙基-5-苯基-2,4,6-(1H,3H,5H)嘧啶叁酮,是5位被乙基和苯基副代替的丙二酰脲衍生物。鉴于PB具拥有镇静、催眠等特殊的生物学干用,在栽物消费中日被用干促长饲料添加以剂[1]。但PB存放在致畸、致癌等毒性干用,其残剩严重为害人的强大健,世界各国均严禁PB用于食品栽物[2]。PB残剩避免疫学检测方法以敏捷、正确、快快、特异等优点顶替了传统的理募化检成方法[3],道德国Dade Behring公司已开收回相干产品,国际尚不见报道。PB半抗原和人工避免疫原的分松及低价、敏感、特异抗体的得到是确立避免疫学检测方法的基础,本试验经度过上述情节的切磋,旨在为PB残剩避免疫学检测方法的确立奠定基础。

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