摘要
目的:筛选出替代血型B抗原的模拟多肽,用多肽抗原替代糖类抗原。方法:抗血型B抗原的单克隆抗体作为固相筛选靶分子,对随机十二肽噬菌体展示文库进行生物淘选(bio-panning),经包被-结合-洗脱-扩增等循环3轮,对筛选的克隆ELISA鉴定,并通过剂量依赖实验验证其结合特异性。最后提取DNA测序,确定模拟肽氨基酸序列。结果:3轮筛选结束,得到2个亲和力较强的十二肽序列TKNMLSL-PVGPG和HSLKHTQMSYSS。结论:经过生物筛选得到模拟多肽序列,利用噬菌体展示技术筛选糖类抗原的模拟肽具有可行性,为糖类抗原的研究提供一种新思路。
OBJECTIVE：To screen peptide mimotopes of blood group B carbohydrate antigen with high affinity for blood group B monoclonal antibody which can replace carbohydrate antigen using a phage display peptide library,and find a new tool for application of blood group B carbohydrate antigen.METHODS：A 12-mer phage peptide library was screened for 3 rounds by using a blood group B monoclonal antibody as target protein according to such a procedure as ＂adsorbing,eluting and amplification＂,and positive clones were selected randomly,confirmed by sandwich ELISA,single strand DNA was extracted from these positive clones and sequenced,and the mimic peptides were deduced by the DNA sequence.RESULTS： After 3 rounds of effective bio-panning,two major mimic peptides with high affinity for target protein were obtained,one peptide sequence was TKNMLSLPVGPG,the other one was HSLKHTQMSYSS.CONCLUSION：The results shows that the motif identified through a 12-mer phage display peptide library can be mimiced and may be a substitute for blood group B antigen.
出处
《中华肿瘤防治杂志》
CAS
2011年第2期95-97,111,共4页
Chinese Journal of Cancer Prevention and Treatment
关键词
二肽类
细菌噬菌体
血型抗原/免疫学
酶联免疫吸附测定
肽库
dipeptides
bacteriophages
blood group antigens/immuneologic
enzyme linked immuneosorbent assay
peptide library
