催化杀虫剂西维因水解的抗体酶制备(Ⅰ)抗原的分子设计、合成与优选

以杀虫剂西维因酯解反应中间过渡态的结构信息进行分子设计，合成了α－萘基磷酸氢酯半抗原，将其与载体蛋白牛血清白蛋白（ＢＳＡ）和鸡卵清蛋白（ＣＥＡ）偶联，制得了具有不同抗原价（４～２２）的合成抗原．经小鼠免疫学实验，优选出了可产生滴度高、特异性强之抗血清的免疫用抗原（Ｈａｐ－ＢＳＡ）．经酶联免疫吸附试验（ＥＬＩＳＡ）测定，用该免疫原制得的鼠抗血清滴度可达１∶３２０００．优选抗原Ｈａｐ－ＢＳＡ可被用作产生催化西维因水解之抗体酶的免疫原．     

催化杀虫剂西维因水解的抗体酶制备(Ⅱ)多克隆催化抗体的制备、纯化及检测

以优选的合成抗原（Ｈ(ａｐ)－ＢＳＡ）免疫动物产生了滴度为１：５５０００（１号兔,Ｒｔ－１）的兔抗多克隆催化抗体．采用盐析和ＰｒｏｔｅｉｎＡ－ＳｅｐｈａｘｏｓｅＣＬ４Ｂ亲和层析法,从兔抗血清中分离出了具有电泳纯度的多克隆催化抗体．经ＳＤＳ－ＰＡＧＥ检测,ＩｇＧｓ相对分子质量为１４８０００．纯化后的多克隆抗体,经ＥＬＩＳＡ法测定,其最低检出活性为２×１０(－６)～４×１０(－６)ｇ／Ｌ．     

从免疫学角度解析抗体酶

抗体酶是具有催化性质的抗体,它同时具备抗体和酶的特征,它的发现是酶学史上的一个重大事件。为帮助生命科学系的学生更好地懂得抗体酶,本文对抗体酶的研究开发思路和历史、制备方法及应用进行了解析。     

Hydrolysis of carboxylate ester catalyzed by a new artificial abzyme based on molecularly imprinted polymer

A new kind of artificial abzyme MIP-3, which contains N-phenyl benzonic amide group and “nanochannel” formed by embedded ZnO nano materials, and is imprinted by a transition-state analogue of p-nitrophenyl methyphosphonate in the hydrolysis of p-nitrophenyl acetate, was prepared by radical co-polymerization. Hydrolytic kinetics of p-nitrophenyl carboxylate catalyzed by MIP-3 was investigated. The results showed that the artificial abzyme exhibited notable substructure selectivity and strong catalytic ability in hydrolysis of p-nitrophenyl acetate