溴苯腈单克隆抗体的制备纯化与性能考察  

Production,Purification and Identification of Bromoxynil Monaclonal Antibody

作　　者：朱江[1] 陆贻通[1] 程国华[1] 

机构地区：[1]上海交通大学农业环境生态研究所,上海201101

出　　处：《农业环境科学学报》2006年第2期527-530,共4页Journal of Agro-Environment Science

基　　金：国家自然科学基金资助项目(20337010);国家基础研究重大项目(2004CB418503);上海市基础研究重点项目(04JC14051)

摘　　要：以人工合成溴苯腈免疫原免疫BALB/CF1小鼠,采用杂交瘤技术制备了3株能稳定分泌抗溴苯腈单克隆抗体的杂交瘤细胞株。用培养扩增后的各建株细胞诱导小鼠生长腹水,腹水效价达10-6～10-7。琼脂双扩试验表明,各单抗蛋白均为IgG,抗体蛋白10B6为IgG1亚类蛋白,3D6和8H2均为IgG2a型亚类蛋白。用辛酸-硫酸铵沉淀法纯化单克隆抗体腹水,对纯化的单抗进一步做交叉反应试验,与其他苯腈类农药交叉反应率低于10%,结果表明该单抗有较好的特异性,可用于对溴苯腈的酶联免疫检测,线性检测范围为10～1000μg·kg-1,溴苯腈检测限为10μg·kg-1。Three hybridoma cell lines secreting anti- bromoxynil McAbs were obtained by immunization of BALB/C F1 mice with the conjugates of bromoxynil immunogen, hybridoma technique and McAbs. 10B6 anti-bromoxynil McAb was categorized into IgG1 subtypes. 3D6 and 8H2 anti- bromoxynil McAb were categorized into IgG2a subtypes. These hybridomas were prepared ascites of the mice, and the titers of the McAbs in the ascites ranged from 10^-6 to 10^-7. McAbs in the ascites were purified by caprylic acid ammonium sulphate precipitation method. The analysis experiment of bromoxynil showed that the cross reactivity were below 10% respectively. So McAbs were therefore considered interesting for developing a bromoxynil -specific ELISA. The quantitative working range of the ELISA was from 10μg·kg^-1 to 1 000μg·kg^-1 with a LOD of 10μg·kg^-1.

关 键 词：溴苯腈 单克隆抗体 杂交瘤技术 酶联免疫吸附测定法 

分 类 号：X830.2[环境科学与工程—环境工程]