摘要
以自行合成的己烯雌酚 (DES)免疫原免疫BALB/c小鼠 ,采用细胞融合、显微克隆等单克隆抗体技术 ,制备了 5株抗DES单克隆抗体分泌杂交瘤细胞株 (7B6、4B10、1G10、4E4、2C7) ;选择其中的 7B6株抗体 ,建立了间接竞争ELISA法 (indirectcompetitiveinhibitionELISA) ,通过初步优化 ,其检测限为 10 pg/孔 ,与同类二苯乙烯类雌激素的交叉反应高 ,而与天然雌激素雌二醇几乎无交叉反应性。这项研究为DES残留检测试剂盒的开发打下了基础。
Four female BALB/c mice aged 4-weeks-old were immunized by intraperitoneal injection with emulsion of DES-MCPE-BSA and Freund’s adjuvant. Splenocytes from the immunized mouse which exhibited the highest titer (1∶12 800) were fused with SP2/0 myeloma cells. The hybridoma cells secreting antibodies against DES were chosen and cloned. Ultimately, five kinds of hybridoma cells(7B6,1G10,4B10,4E4 and 2C7) were cultured. A method of indirect competitive inhibition ELISA was developed by using the supernatant of 7B6 hybridoma cells . The calibration curve in the method was y= -31.67x+95.892(r=0.983)in 1-250 ng/mL. The half maximal inhibition to Ag-Ab reaction was 500 pg per well or so,LOD= 10 pg per well. Cross reactivityof 7B6 antibody with HEX,DIEN,DES-MG was rather high(1-50 ng/mL) respectively, but with E 2 was considerablyfew.
出处
《中国兽医科技》
CAS
CSCD
北大核心
2003年第11期12-16,共5页
Chinese Journal of Veterinary Science and Technology
基金
国家科技攻关计划项目畜禽规模化养殖项目子课题 :兽药安全评价与残留检测技术(2 0 0 2BA5 14A 14 0 3)