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目的 :制备新化合物 1 羟基 1 对胺基苯基甲磷酸单频哪基醇酯 (P4 )的单克隆抗体 (mAb)。方法 :以P4为半抗原 ,与血蓝蛋白 (LPH)化学偶联制备人工抗原 (P4 LPH) ,并以其免疫BALB/c小鼠 ,制备抗P4的mAb。用竞争抑制酶免疫分析法 ,测定mAb对梭曼等 7种配体的结合活性 ,结果以mAb与P4 BSA的结合被抑制 5 0 %时的配体浓度 (IC50 ) ,表示mAb的交叉反应性。结果 :建立了 9株对P4 BSA阳性反应的mAb ,它们分属 3种Ig亚类 ;6株 (2B2、3A10、3D5、4F1、6C9和 6H4 )为IgG1,1株 (3B9)为IgG2b ,2株 (1B3和 6H5 )为IgM。 9株mAb中 ,8株具有与半抗原P4的结合的特异性。4株 (1B3、2B2、3D5和 4F1)具有梭曼结合活性 ,其IC50 值分别为 10 -3 、10 -3 、10 -5和 10 -6mol/L ;1株 (4F1)具有与对氧磷结合的特异性 ,其IC50 值为 10 -5mol/L。 4株腹水的mAb鼠滴度高 ,分别达 10 -6(1B3和 2B2 )和 10 -8(3D5和 4F1)。结论 :用新的半抗原P4成功地制得抗梭曼和抗对氧磷的mAb ,可用于梭曼抗体酶、梭曼的免疫抗毒和免疫检测研究
OBJECTIVE: To prepare a monoclonal antibody (mAb) for the novel compound 1 hydroxy 1-p-aminophenylphosphates mono-pinacol ester (P4). Methods: The anti-P4 mAb was prepared by immunizing BALB / c mice by chemically conjugating P4 LPH with P4 LPH and chemically coupling with LPH. The binding activity of mAb to seven ligands such as soman was measured by competitive inhibition enzyme immunoassay. As a result, the ligand concentration (IC50) when the binding of mAb to P4 BSA was inhibited at 50% was measured, indicating the cross-reactivity of the mAb . Results: Nine mAbs positive for P4 BSA were established, which belonged to three Ig subclasses; 6 (2B2, 3A10, 3D5, 4F1, 6C9 and 6H4) were IgG1 and 1 strain (3B9) Strains (1B3 and 6H5) are IgM. Of the 9 mAbs, 8 strains had specificity for binding to hapten P4. The 4 strains (1B3, 2B2, 3D5 and 4F1) had the binding activity of soman with IC50 values of 10 -3, 10 -3, 10 -5 and 10 -6 mol / L respectively. One strain (4F1) The specificity of the binding is 10 -5 mol / L. The 4 ascitic mAb mice had high titers of 10 -6 (1B3 and 2B2) and 10 -8 (3D5 and 4F1), respectively. Conclusion: The mAb against soman and anti-paraoxon can be successfully prepared with the new hapten P4, which can be used in the immune anti-virus and immunoassay of soman antibody and soman