己烷雌酚免疫原的合成及其鼠源抗血清的制备  

Synthesis of immunogen and preparation of mouse antiserum against Hexestrol

作　　者：杨兴东 段宇莹 李琳 YANG Xing-dong;DUAN Yu-ying;LI Lin(Institute of Food and Drug Inspection,Zhoukou Normal University, Zhoukou 466001,Henan Province,China)

机构地区：[1]周口师范学院食品药品检验检测中心

出　　处：《中国生物制品学杂志》2019年第7期798-802,共5页Chinese Journal of Biologicals

基　　金：河南省自然科学基金(182300410093);河南省科技攻关项目(182102310683)

摘　　要：目的合成己烷雌酚(hexestrol,HES)免疫原,并制备其鼠源多抗血清。方法在HES的适当位点上引入活性基团羧基(-COOH)得到HES的半抗原,采用EDC/NHS法合成人工免疫原HES-BSA和检测抗原HES-OVA,经紫外扫描和SDS-PAGE鉴定HES与载体蛋白BSA、OVA是否成功偶联;用免疫原(HES-BSA)免疫3只BALB/c小鼠,获得抗HES多抗血清(polyclonal antibody serum,pAbs),ELISA法检测其效价,间接竞争ELISA法检测其敏感性,交叉反应(cross-reactivity,CR)试验检测其特异性。结果免疫的1号小鼠pAbs效价达到1.28×10^4,2号和3号小鼠均在6.4×10^3以上;1号小鼠HES pAbs对HES的50%抑制浓度(half maximal inhibitory concentration,IC50)为19.93 ng/mL,除与己烯雌酚有11.63%的CR外,与其他结构类似物无CR。结论获得了效价高及敏感、特异的多抗血清,为进一步的免疫学快速检测提供了技术支持。Objective To synthesize the immunogen against Hexestrol (HES) and prepare its polyclonal antiserum. Methods The hapten against HES was obtained by introduction of reactive carboxyl groups(-COOH)into the appropriate site of HES,based on which the artificial immunogen HES-BSA and detection antigen HEB-BSA were synthesized by EDC / NHS method. The successful couplings of HES and carrier proteins (BSA and OVA)were identified by ultraviolet scanning and SDS-PAGE. Three BALB / c mice were immunized with immunogen HES-BSA,and the prepared polyclonal antibody serum(pAbs)against HES were determined for titer by ELISA,and for specificity by cross-reactivity(CR)test. Results The titer of pAbs induced in mouse 1 reached 1. 28 × 10^4,while those in mice 2 and 3 reached more than 6. 4 × 10^3. The half maximal inhibitory concentration (IC50 )of pAbs against HES in mouse 1 was 19. 93 ng / mL. The pAbs showed 11. 63% cross-reactivity(CR)with ethylstilbestrol,while showed no CR with other structural analogues.

关 键 词：己烷雌酚 免疫原 抗血清 ELISA 

分 类 号：S852.4[农业科学—基础兽医学]