甲砜霉素及氟苯尼考胺单克隆抗体的制备及特性鉴定

将氟苯尼考胺与HSA载体蛋白相连作为抗原免疫Balb/c小鼠,经杂交瘤技术获得了两株能稳定分泌抗甲砜霉素单克隆抗体的杂交瘤细胞株1E及5C。用间接竞争ELISA方法测定,其细胞株1E单克隆抗体的细胞上清液效价为1∶5×103,腹水效价为1∶1×106,抗体亚类为IgG2b,50%抑制浓度(IC50)为15μg/L。其分泌的单克隆抗体与其结构类似物氟苯尼考及氟苯尼考胺的交叉反应率为10.4%和10.8%,和其他化合物基本无交叉反应。体外传代培养和冻存复苏后抗体分泌稳定,可为检测试剂盒提供长期稳定的抗体。

Preparation and Identification of Monoclonal Antibody Against Thiamphenicol and Florfenicol-amine

Balb/c mice were immunized by artificial antigen florfenicolamine- HSA, the hybridoma lines 1E and 5C that secreted anti -thiamphenicol mAb were established by using monoclonal antibody hybridoma technology. The titers that the hybridoma 1E secreted monoclonal antibody in cell line supernatant and ascites were 1：5 ×10^3 and 1 ： 1 × 10^6 by indirect ELISA respectively. The isotypes in ascites was IgG2b, IC50 was about 15μg/L. Cross reactivity to florfenicol and florfenicolamine were 10.4% and 10.8% respectively, little cross reactivity to other compounds. Monoclonal antibodies were stable secreted in vitro cell culture and cryoprotectant. Long -term stability of antibody which kit used can be provided.