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溴苯腈单克隆抗体的制备纯化与性能考察

Keywords: 溴苯腈,单克隆抗体,杂交瘤技术,酶联免疫吸附测定法

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Abstract:

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。

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