摘要
目的对抗氯霉素单克隆抗体4D10进行纯化与特性鉴定,以获得可用于检测氯霉素残留试剂盒的单克隆抗体。方法用两步硫酸铵法和G蛋白亲和层析法纯化单克隆抗体,用间接酶联免疫吸附试验、竞争ELISA等方法测定抗体效价、特异性、敏感性与亲和性以及抗体与其它氯霉素结构类似物的交叉反应率。结果纯化后抗体的纯度高达95%;对抗体进行相关性质的鉴定得出：4D10效价为1∶2.56×105,其亲和力常数可达2×108M-1,该抗体与CAP-BSA,CAP-OVA有反应,与BSA、OVA无反应;与氯霉素琥珀酸钠、氯霉素反应,而与其它氯霉素结构类似物无交叉反应。结论抗氯霉素单克隆抗体4D10效价高、特异性强、亲和力高,可利用该抗体制备检测氯霉素残留的ELISA试剂盒。
Objective To prepare and to characterize the monoclonal antibody （McAb） against chloramphenicol （CAP）. The McAb was used for the kit of detecting chloramphenicol residues.Methods Two-step ammonium sulfate precipitation and protein G purification by affinity chromatography were used to purify McAb. Competitive inhibition enzyme linked immunosorbent assay （ciELISA） was used to determine the antibody titer and competitive ELISA was used to determine specificity,sensitivity,affinity,and the cross-reactivity between the antibody and chloramphenicol analogs.Results The purity of the antibody was 95%,the titer of McAb 4D10 was 1：2.56×105,its affinity was 2×108 M-1,and it had no reactivity with BSA,OVA and the chloramphenicol analog.Conclusion McAb 4D10 had high titer,high specificity,and high affinity,and could be used to develop CAP immunoassay kits for detection of CAP residues.
出处
《广东药学院学报》
CAS
2010年第4期420-422,共3页
Academic Journal of Guangdong College of Pharmacy
基金
广东省科技攻关计划资助项目(2006B20501005)
广州市科技计划资助项目(2006JI-C0221)
广东省医学科学技术研究基金(A2006323)
关键词
氯霉素
单克隆抗体
亲和常数
chloramphenicol （CAP）
monoclonal antibody
affinity
