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。
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 ×10^4 , respectively. A competive ELISA test was developed with anti-KAN monoclonal antibody and the sensibivity of 0.5 ng/mL,the ICs0 of 3.7 ng/mL. The antibody against kanamycin was selective toward tobra- mycin 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.
出处
《华北农学报》
CSCD
北大核心
2010年第2期121-124,共4页
Acta Agriculturae Boreali-Sinica
基金
天津市科委基础研究项目(07JCYBJ11800)
