基于磺胺类药物(Sulfonamides,SAs)公共母环(对氨基苯磺酰胺)结构,合成了两种具有该结构的半抗原:2-(对氨基苯磺酸氨基)一4一噻唑乙酸(Ts)和对氨基苯磺酰胺基苯甲酸(SS)。采用碳二亚胺法合成了免疫原和包被原,分别免疫10只新西兰大白兔,制备广谱性磺胺类药物抗体。应用间接竞争酶联免疫吸附分析(ELISA)检测血清的灵敏度和特异性,发现所有兔子对TS~BSA和SS-BSA均产生较高效价和特异性。其中SS-1号和TS-6号兔的血清灵敏度最好,效价分别为1:80000和1:160000,对10种磺胺类药物均有不同程度的识别能力。SS-BSA组IC50为257.9~6590,9ng/mL,而TS-BSA组IC50为12.9~586.1ng/mL。结果表明,具有磺胺母环结构的半抗原TS和SS,可以应用于广谱性磺胺类抗体的制备。通过10只兔子免疫试验证明,应用SS获得的血清具有较高的簇特异性、相似的亲和性,优于TS。
Hapten TS and SS with the common structure(p-anilinesulfonamide) of sulfonamides were synthesized. Haptens were conjugated to bovine serum protein and albumin by EDC method, which were used to immunize 10 New Zealand White rabbits to make the broad-specificity polyclonal antibody and coating antigen, respectively. The sensitivity and specificity of antisera were measured by ELISA. The results show that all rabbits presented high antisera titer and relatively specific recognition to sulfonamides. The sensitivity of the antisera of No. SS-1 and TS-6 to sulfonamides in ELISA were the best two with the antisera titer of 1 : 80 000 and 1 : 160 000, respectively. The antisera of No. SS-1 could recognize 10 sulfonamides showing 50% inhibition(IC50 ) at a concentration of 257.9 ng/mL to 6 590.9 ng/mL, while the antisera of No. TS-6 could at 12.9 ng/mL to 586.1 ng/mL, respectively.