氯霉素单克隆抗体制备以及化学发光检测试剂盒的研制

Preparation of chloramphenicol monoclonal antibodies and chemiluminescence immune assay kit

通过用钯炭催化氯霉素,通入氢气,制备出氯霉素半抗原,通过动物免疫、细胞融合,获得杂交瘤细胞株,再由杂交瘤细胞株分泌得到抗氯霉素单克隆抗体,从而制备出能够检测鱼肉和虾肉样本中氯霉素药物的化学发光试剂盒。试剂盒对鱼肉的检测限为0.047μg/kg,对虾肉的检测限为0.045μg/kg,对鱼肉的回收率为83.5%～109.3%,批内、批间相对标准偏差(RSD)范围均<10%。氯霉素单克隆抗体与棕榈氯霉素、甲砜霉素、氟甲砜霉素、氟苯尼考胺的交叉反应率均<1%。试剂盒能够在4℃保存1年。

The chloramphenicol hapten was prepared by a sequence of reactions that using chloramphenicol,palladium charcoal and hydrogen.Chloramphenicol monoclonal antibodies were prepared by animal immune,cell fusion,hybridoma screening and other steps,and then the chemiluminescence immune kit that was capable of detecting chloramphenicol in fish and shrimp was prepared.The detection limits of the kit for fish and for shrimp were 0.047μg/kg and 0.045μg/kg,respectively.The recoveries for fish were between 83.5%-109.3%.The relative standard deviation(RSD)was less than 10%.The cross reaction rate with palm chloramphenicol,thiamphenicol,florfenicol and florfenicol amine was less than 1%.The kit could be kept at 4℃for one year.