黄曲霉毒素B1抗独特型抗体的制备和应用研究I抗独特型抗体的制备和性质研究

以黄曲霉毒素B1(AFB1)与牛血清蛋白（BSA)的连接物AFB1-BSA注射免疫兔子获得抗AFB1抗血清，经(NH4)2SO4沉淀、酶切处理与亲和层析分离纯化后，得到抗AFB1独特型抗体Ab1及其酶切片段Fab1，然后再将Fab1注射免疫BALB／c小鼠，得到抗(抗AFB1)独特型抗体Ab2及其酶切Fab2。研究了Ab2和Fab2的性质，结果表明，Ab2和其Fab2部仅与Ab1及其Fab1反应，而不和抗桔霉素等其他抗体反应，有较好的特异性。以AFB1与卵清蛋白(OV)的连接物AFB1-OV为包被抗原，Fab1为反应抗体，Ab2和Fab2为竞争抗原，达到50％的竞争抑制率时，Ab2和Fab2的浓度分别为3．98ug／ml和1．12ug／ml；而以Ab2和Fab2为包被抗原，Fab1为反应抗体，AFB1为竞争抗原，达到50％的竞争抑制率时，AFB1的浓度分别为44．67ug/L和6.31ug／L，这表明无论是Ab2还是Fab2都和AFB1有很好的内影像关系，可以作为AFB1的替代品用于AFB1的免疫学检测。但是相对而言，由于Fab2的分子量小，反应时的空间位阻小，所以Fab2更适合于用作AFB1的替代品。

PREPARATION AND APPLICATION OF AFLATOXIN B1(AFB1) ANTI-IDIOTYPE ANTIBODY I.STUDY ON PREPARATION AND CHARACTERIZATION OF AFB1 ANTI-IDIOTYPE ANTIBODY

Anti-AFB1 antiseria were produced with the AFB1-BSA conjugate injecting rabbits, and idiotype antibody (Ab1) and Fab1 were achieved after (NH4)2SO4 precipitation, enzyme hydrolyzation treatment and affinity chromatography. BALB/c mice were immunized with Fab1, and anti-idiotype antibody (Ab2) and Fab2 were gotten. Characterization of Ab2 and Fab2 was investigated. The results revealed that Ab2 and Fab2 had the high specificity, and they could only react with Ab1 and Fab1, whereas not react with the other antibodies such as anti-citrinin antibody. When AFB1-OV was used as coating antigen, Fab1 as reaction antibody, and Ab2 and Fab2 as competitive antigens, the concentration of Ab2 and Fab2 was 3.98ug/ml and 1.12ug/ml at 50% competitive inhibitory ratio, respectively. When Ab2 and Fab2 were used as coating antigens, Fab1 as reaction antibody, and AFB1 as competitive antigen, the AFB1 concentration was 44.67ug/L and 6.31ug/L at 50% competitive inhibitory ratio, respectively. These results showed that there was internal image between AFB1 and Ab2 (or Fab2), so both Ab2 and Fab2 which could substitute AFB1, might be used in AFB1immunological analysis. However, comparatively, Fab2 with lower molecular weight, was fitter to replacing AFB1 due to less steric hindrance.