摘要
采用重氮化法将磺胺甲噁唑(SMZ)与人血清白蛋白偶联形成免疫原,免疫大白兔制备多克隆抗体。通过紫外扫描和高效液相色谱分析偶联物,免疫抗原的偶联率为19∶1,改良辛酸-饱和硫酸铵纯化抗体,多克隆抗体效价达1∶105,间接酶联免疫吸附试验建立的SMZ检测的线性范围在0.1ng/mL^10μg/mL之间,50%抑制率检测限为22.53ng/mL,抗体与磺胺甲噁唑交叉反应率为100%,与其他7种磺胺药交叉反应率很低。表明制备的磺胺甲噁唑多克隆抗体具有很高的特异性,可用于SMZ在动物性产品中残留的检测。
A polyclonal antibody （pAb） specific for sulfamethoxazole （SMZ） was prepared. Hapten SMZ was coupled with carrier protein human serum albumin （HSA） to form an antigen SMZ-HSA by diazotization method. The conjugates used to immunize New Zealand white rabbit were identified with UV scanning spectrum and high performance liquid chromatography （HPLC） and the couple rate of the antigen was 19 ： 1. After the antisera were obtained and purified using a modified n-caprylic acid-saturated ammonium sulfate method, the specificity of pAb was detected by indirect enzyme-linked immunosorbent assay （ELISA）. It showed that the titer of pAb was 1 ： 10^5 and the detection limit in term of 50% inhibition rate was 22.53 ng/mL in the linear range of 0.1 ng/mL～10 μg/mL. The cross-reaction rate between SMZ and the pAb was 100% and the cross-reaction was very lower between the pAb and seven kinds of sulfonamides. The results showed that the anti-SMZ pAb had a high specificity and was good enough for immunoassay of SMZ in edible animal products.
出处
《动物医学进展》
CSCD
2006年第1期72-75,共4页
Progress In Veterinary Medicine
基金
北京市农委资助(20020103)
北京市教委资助(200310020084)
北京市组织部资助(20041D0502106)
关键词
磺胺甲噁唑
多克隆抗体
酶联免疫吸附试验
sulfamethoxazole （SMZ）
polyclonal antibody （pAb）
enzyme-linked immunosorbent assay （ELISA）
