摘要： 采用碳二亚胺(EDC)法合成卡那霉素完全抗原,免疫BALB/c小鼠,通过杂交瘤技术获得3株能稳定分泌抗卡那霉素单克隆抗体的杂交瘤(5D7、6H8、7G4)。选择其中的5D7株抗体,经ELISA鉴定,细胞上清的效价达10³以上,腹水效价达到5×10⁴以上;建立了间接竞争ELISA法,通过优化,其检测限为0.5ng/mL,抗体IC₅₀为3.7ng/mL,与妥布霉素交叉反应率为63.8%,与其他抗生素无交叉。可见ELISA具有快速、敏感、特异、简便等特点,适合于KAN残留的快速检测,具有较高的推广应用价值。

关键词: 卡那霉素, 单克隆抗体, ELISA

Abstract: The antigen of kanamycin synthesized by EDC reaction was used for preparation of a hybridoma cell line.which secretes monoclonal antibodies against kanamycin.The titers of antibody in cell supematant and ascites were 1∶103 and 1∶5×104,respectively.A competive ELISA test was developed with anti-KAN monoclonal antibody and the sensibivity of 0.5 ng/mL,the IC50 of 3.7 ng/mL.The antibody against kanamycin was selective toward tobramycin with cross-reactivity of about 63.8%,and,wasn't detective cross-reactivity toward others.The ELISA rapidity,sensitivity,specificity and briefness and was proved to be used in the rapid test of KAN residues in animal food.

Key words: Kanamycin, Monoclonal antibody, EL ISA

中图分类号: 

• Q813.2