摘要
目的 从构建的A型流感病毒核蛋白(Nucleoprotein)噬菌体单链抗体库中筛选阳性克隆,在大肠杆菌HB2151中表达流感病毒核蛋白噬菌体单链抗体,为研制A型流感病毒免疫渗漏试剂盒建立基础。 方法 经过连续三轮固相"亲和-洗脱-富集"筛选,丰富噬菌体抗体库中阳性克隆的比例,采用ELISA和SDS-PAGE方法对大肠杆菌培养上清液和细菌周质腔蛋白进行分析目的产物。 结果 随着淘洗轮数的增加,NP抗原特异性的噬菌体抗体得到了高度富集,每轮淘洗可以增加10倍左右的特异性的噬菌体抗体;从96个克隆中筛选到10个阳性克隆,其中三个阳性信号较强,分别为A1、E1和G3。其OD450nm分别达到0.469、0.582和0.507。 结论 利用噬菌体抗体库技术,初步在大肠杆菌中表达了A型流感病毒噬体单链抗体。
Objective To express the phage single chain antibody against nucleoprotein of influenza virus type A in E.coli strain HR2151 by screening the positive clone from the phage antibody library against nucleoprotein, and prepare for the construction of fast leak kit of Influenza virus type A. Methods The ratio of positive clone in the phage antibody library was enriched by three-round continual solid panning, ELISA and SDS-PAGE methods were used to analyze the protein secreted in the supernatant and periplasmic. Results Along with the increase of rounds nucleoprotein specific phage antibody was highly enriched and the screening efficiency was increased 10 folds than the former round.Ten clones were screened from 96 clones and among the 10, there were 3 stronger positives with OD450mm value of 0.469,0.582 and 0.507, respectively. Conclusion By using phage chain antibody library technique, phage chain antibody library technique, phage single antibody against nucleoprotein of influenza virus type A was preliminary expressed in E. coli.
出处
《中国热带医学》
CAS
2005年第1期19-21,24,共4页
China Tropical Medicine
