摘要
为建立氯唑西林(cloxacillin,CLOX)残留的酶联免疫(enzyme-linked immuno sorbent assay,ELISA)检测方法,试验采用碳化二亚胺法,将氯唑西林偶联于载体蛋白牛清蛋白(Bovine albumin,BSA)和卵清蛋白(ovalbumin,OVA),制得免疫抗原(CLOX-BSA)和包被抗原(CLOX-OVA),并通过红外光谱扫描(infrared spectroscopy,IR)鉴定表明CLOX人工抗原合成成功。用免疫抗原免疫5只小鼠,制备CLOX多克隆抗体,通过间接ELISA法测定其对CLOX的效价及特异性。结果表明:用CLOX-BSA免疫制备的抗体效价达1∶64000以上,特异性较强,对CLOX的半抑制质量浓度IC50值为(5.64±0.03)ng/mL,IC15值为(22.43±0.02)ng/mL。说明试验得到了灵敏度高、特异性强及具有开发性的抗体,可为食品安全检验试剂盒的开发提供基础。
A method for detecting residual enzyme-linked immunosorbent(ELISA)of cloxacillin(CLOX)was established.In the experiment,CLOX-BSA and CLOX-OVA were prepared by coupling cloxacillin to carrier protein bovine albumin(BSA)and ovalbumin(OVA)by carbodiimide method,and the synthesis of CLOX-BSA and CLOX-OVA was proved to be successful by IR scanning.KM mice were immunized with immunizing antigen to prepare CLOX polyclonal antibody,and its titer and specificity for CLOX were determined by indirect ELISA.The results showed that the preparation with CLOX-BSA immune antibody titer for 1∶64000 above,strong specificity,quality of CLOX half inhibition concentration of(5.64±0.03)ng/mL,and the IC15 value was (22.43±0.02) ng/mL.It showed that the experiments had obtained high sensitivity, specificity and developmental antibodies, which could provide a basis for the development of food safety testing kits.
作者
张冬昊
赵桐桐
何扩
ZHANG Dong-hao;ZHAO Tong-tong;HE Kuo(Key Laboratory of Analysis and Testing of Agricultural Products Food Quality and Safety in Hebei Province,Hebei North University,Zhangjiakou 075000,Hebei,China)
出处
《食品研究与开发》
CAS
北大核心
2020年第4期106-111,共6页
Food Research and Development
关键词
氯唑西林
偶联
免疫
人工抗原
抗体
特异性鉴定
cloxacillin
coupling
immune
artificial antigen
antibody
specific identification
