论文部分内容阅读
以1,4-丁二醇二缩水甘油醚(双环氧试剂)为偶联剂,合成桔霉素-蛋白偶联抗原CIT-BSA,将偶联抗原免疫BALB/C小鼠,取脾细胞与小鼠骨髓瘤细胞SP2/0进行融合,应用有限稀释法进行筛选,经过克隆化后筛选到一株稳定分泌抗桔霉素抗体的杂交瘤细胞株H2-F8.该单克隆抗体经过初步鉴定,抗体类型为IgM类,抗体的相对亲和力为4.17×108L/mol,单抗与黄曲霉毒素B1、赭曲霉毒素A、脱氧雪腐镰刀菌烯醇和玉米赤霉烯酮等毒素的交叉反应率均低于0.1%,与红曲色素中的橙色素和红色素的交叉反应率均低于0.01%.在此基础上建立了间接竞争ELISA检测方法,线性范围为0.05～1.0μg/L,IC50值为0.3μg/L.结果为快速检测桔霉素的酶联免疫检测方法的建立和检测试剂盒的研制提供技术依据.
CIT-BSA was synthesized with 1, 4-butanediol diglycidyl ether (double epoxy reagent) as the coupling agent, BALB / C mice were immunized with conjugated antigen and splenocytes And mouse myeloma cells SP2 / 0 fusion, limited dilution method for screening, cloned and screened to a stable secretion of anti-citrinin antibody hybridoma cell line H2-F8. The monoclonal antibody was initially identified , And the antibody type was IgM. The relative affinity of the antibody was 4.17 × 108 L / mol. The cross-reactivity of McAb with aflatoxins B1, ochratoxin A, deoxynivalenol and zearalenone Less than 0.1%, and the cross-reaction rate between red pigment and orange pigment was lower than 0.01% .And indirect competitive ELISA was established on the basis of this method.The linear range was 0.05-1.0μg / L, the IC50 value Was 0.3μg / L. Results For the rapid detection of citrinin enzyme-linked immunosorbent assay and the establishment of test kit provide a technical basis.