应用噬菌体展示技术筛选氯胺酮模拟表位的研究

Research on utilization of phage display technology to screen chloramine ketone simulated surface point

目的应用噬菌体展示技术筛选氯胺酮模拟表位,为建立无毒ELISA检测体系提供基础。方法以抗氯胺酮单克隆抗体为配体,在噬菌体七肽库中淘选氯胺酮的模拟表位。经4轮淘选后,选择可与氯胺酮单克隆抗体有不同程度结合的噬菌体,并进一步以间接竞争ELISA确定与盐酸氯胺酮存在竞争抑制的阳性克隆。结果发现10个噬菌体均可与氯胺酮单克隆抗体不同程度的结合;以间接竞争ELISA确定其中5个阳性克隆与盐酸氯胺酮存在竞争抑制现象;5个阳性克隆测序结果表明,获得了4种不同的氨基酸序列。结论初步判断这4个噬菌体展示了氯胺酮模拟抗原表位。

Objective Utilize phage display technology to screen chloramine ketone simulated surface point,providing the basis for establishing drug-free ELISA test system.Methods Use the anti chloramine ketone monoclonal antibody as the ligand,and screen the Ph.D.-7TM phage display peptide library to select the chloramine ketone simulated surface point.After 4 rounds of screening,select the phase,which can combine the chloramines ketone monoclonal antibody with different degrees,and further,determine the positive clone with competitive inhibition of the hydrochloric chloramine ketone by the indirect competitive ELISA method.Results It is discovered that all the 10 phages can combine the chloramine ketone monoclonal antibody with different degrees.Use the indirect competitive ELISA method to determine,the phenomenon that 5 positive clones,among them,have competitive inhibition with the hydrochloric chloramine ketone exists.The sequencing results of those 5 positive clones show that 4 different amino acid sequenced are obtained.Conclusion Tentatively judge these 4 phages display the chloramine ketone simulated antigen surface points.