摘要
目的用针对HSV-2gD的单克隆抗体(McAb)从噬菌体展示随机12肽库中筛选HSV-2gD基因的抗原模拟表位,寻找更有效的小片段短肽作为HSV新型基因工程疫苗的候选抗原。方法利用HSVgDMcAb淘筛噬菌体随机12肽库,经"吸附-洗脱-扩增"的4轮筛选,随机挑取噬菌体克隆经酶联免疫吸附实验(ELISA)进行鉴定,并对阳性克隆所携带的外源DNA片段进行序列测定和计算机辅助分析。结果经4轮生物淘筛后特异性噬菌体克隆得到了富集,对阳性克隆的分析结果表明P-PLW和PYH--H氨基酸序列是模拟gD基因抗原表位的骨架结构,携有此短肽的噬菌体可以与McAb特异结合。结论从噬菌体随机12肽库中筛选到的外源序列可以模拟HSV-2gDMcAb针对的抗原表位,可能是HSV-2gD一个抗原表位的替代抗原,为进一步研究HSV-2基因疫苗奠定了基础。
[Objective] The aim of this study is to obtain mimic peptide epitopes of HSV-2 glyeoprotein D by screening of a phage display peptide library and to provide a candidate antigen for the preparation of genetic engineering vaccine with more oligo-peptides against type 2 herpes simplex virus. [Methods] A 12-mer phage peptide library was screened for 4 rounds by using a HSV-2 gD monoclonal antibody （McAb） according to such a procedure as ＂adsorbing, eluting and amplification＂, and positive clones were selected randomly, confirmed by ELISA and sequencing, then further analysis was conducted with a certain computer software. [Results] After 4 rounds of effective bio-panning, two major sequences as P-PLW and PYH--H carried in the positive phage clones were confirmed for the mimic epitope, and the clones containing this motif could react with the McAb specifically. [Conclusion] The results showed that the motif identified through a 12-mer phage display peptide library could mimic and may be a substitute for one epitope of HSV-2 gD, which may provide a candidate for DNA vaccine of HSV-2.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2008年第9期1194-1196,共3页
China Journal of Modern Medicine
基金
山东省卫生厅资助项目
(No:H1A1)
