目的:筛选法氏囊病毒(IBDV)单克隆抗体所针对的模拟抗原表位。方法:以纯化的3株IBDV单克隆抗体为靶分子用噬菌体12肽库筛选相应抗原模拟表位;应用间接和竞争ELISA鉴定所筛选的阳性样品;最后对与抗体高亲和结合的噬菌体进行测序分析。结果:经过四轮淘洗,随机挑取30个克隆经间接ELISA鉴定,发现其中22个克隆结合活性较高。进一步应用竞争ELISA,获得14个噬菌体克隆,其抑制率高达50%以上。对这些噬菌体进行DNA测序,并分析比对了IBDV相应序列,确定了3个抗原模拟表位。结论:通过噬菌体随机肽库成功筛选出3个IBDV模拟表位,为进一步研究IBDV抗原性质奠定了基础。
Objective:Screening of mimic epitopes from phage display 12 peptide library by monoclonal antibody against infectious bursal disease virus.Methods:The phage display 12 peptide library was screened with three strains of purified mAb against IBDV as target molecules for the mimic epitopes.The indirect and competitive ELISAs were used for identification of positive samples.Some of phages with high affinity binding antibody were sequenced and analyzed.Results:After forth panning,30 clones were randomly picked and identified by indirect ELISA;among these clones,there were 22 clones binding antibodies with higher affinity.14 phage clones had been obtained from the indirect competitive ELISA,and the inhibition rates reached up to 50%.Sequence analysis of these positive phages showed that 3 amino sequences were the mimic epitopes of IBDV.Conclusion:3 mimic epitopes of IBDV antigen were obtained successfully by the phage display library,which provides bases for the research of IBDV antigen.