摘要
目的:用抗-HSV-2gD单克隆抗体(mAb)从噬菌体随机12肽库中筛选HSV-2gD抗原模拟表位序列.方法:利用HSV-2gDMcAb淘筛噬菌体随机12肽库,经四轮富集筛选,随机挑取阳性克隆扩增后进行ELISA鉴定,同时提取其基因组DNA进行序列测定,并利用生物信息学软件对包膜糖蛋白D(gD)进行B细胞抗原表位分析.结果:经四轮生物淘筛后,特异性噬菌体克隆得到了富集.经过对阳性克隆携带的随机12肽序列进行综合生物信息学分析,得到了PYH-H和P-PLW两个模拟HSV-2gD抗原表位的主要氨基酸基序.结论:用噬菌体随机12肽库成功筛选到了两个模拟HSV-2gD抗原表位的主要氨基酸基序,可模拟McAb针对的抗原表位,可能是HSV的替代抗原.该研究方法为研制更有效及更广谱的HSV基因疫苗提供了依据.
AIM： To screen antigenic mimotope of herpes sim- plex virus type II glycoprotein D （HSV-2gD） by the monoclonal antibodies against HSV-2gD using a 12-mer Phage display peptide library. METHODS： A 12-mer phage peptide library was bio- panned for 4 rounds with the monoclonal antibodies against HSV- 2gD. Specific phage clones of the HSV-2gD were identified by ELISA. ssDNA of positive clones were extracted and purified for sequencing and confirmed by computer assistant analysis. RE- SULTS： After 4 rounds of effective biopanning, we got specific phage clones. Analysis with computer demonstrated that the PYH- H and the P-PLW were the main amino-acid motif. CONCLU- SION： The PYH-H and the P-PLW are mimotope of HSV-2gD and may be a substitute antigen of gD. This strategy provides the potential for preparing more effective epitope vaccine.
出处
《第四军医大学学报》
北大核心
2007年第21期1937-1939,共3页
Journal of the Fourth Military Medical University
基金
山东省卫生厅资助项目(HZ149)
