采用噬菌体展示技术筛选玉米赤霉醇的单链抗体

Selection of anti-zeranol ScFv antibody by phage display technology

目的从噬菌体单链抗体库中筛选玉米赤霉醇(ZER)的阳性克隆。方法人工抗原ZER-BSA作为包被原,采用负筛选,经过连续3轮固相"亲和-洗脱-富集"筛选,富集了噬菌体抗体库中阳性克隆的比例,同时采用Trypsin和ZER竞争洗脱2种洗脱方法。ELISA鉴定阳性克隆,运用SDS-PAGE方法对大肠杆菌分泌蛋白进行了时间条件的摸索。结果随着淘洗轮数的增加,特异性噬菌体抗体得到了高度富集,从6个96孔板的克隆中筛选到10株阳性克隆(阳性大于阴性3倍视为阳性克隆),将菌株转换成功的5株测序发现有4株不同的阳性克隆,对一株进行了原核系统的表达,SDS-PAGE确定了目的条带的存在,相对分子质量为Mr28000。结论为在天然噬菌体抗体库中制备玉米赤霉醇的抗体奠定了基础。

Objective To screen the single chain antibody against zeranol (ZER) from the phage-display antibody library. Methods ELISA plate was coated by BSA-conjugated ZER,while subtractive panning was employed. The ratio of positive clone in the phage antibody library was elevated by three rounds of continual 'adhesion-elution-amplification' solid panning,using two ways of eluting with trypsin and ZER. The positive clones was evaluated by phage ELISA. SDS-PAGE method was used to analyze the best time for getting mor...