生命科学研究2017，Vol.21Issue(1)：16-22,7.DOI:10.16605/j.cnki.1007-7847.2017.01.004
人巨细胞病毒包膜糖蛋白B主要中和表位单克隆抗体的制备
Preparation of Monoclonal Antibodies Against the Main Neutralizing Epitopes of HCMV gB
摘要
Abstract
Human Cytomegalovirus (HCMV) is ubiquitous in humans,and could cause a variety of diseases.It presents a high infection rate worldwide,which poses a serious threat to the health of human beings.Here,main coding sequence containing neutralizing antigenic epitopes AD-1,AD-2 and AD-3 (named AD13) on the HCMV gB was synthesized according to the gene sequence of HCMV AD169.Expression vector pET-32a was digested by EcoR I/Xho I,and the coding sequence of neutralizing antigenic epitopes was ligated to pET-32a vector to construct the recombinant prokaryotic expression plasmid.Western-blot and indirect ELISA were used to investigate the recombinant expression,purity and immunoreactivity of AD13 protein.The BALB/c mice were immunized with the purified protein,and then the monoclonal antibodies (mAbs) against AD13 protein were prepared and screened.SDS-PAGE showed that the relative molecular mass of recombinant AD13 protein was 35 kD,and Western-blot and indirect ELISA results revealed that the recombinant AD13 protein,as an antigen,had significant immunoreactivity.After cell fusion and screening,five hybridoma cell strains secreting monoclonal antibodies against AD13 were obtained,which have specificity in reaction with the AD13 antigen.This provides diagnostic raw materials and research foundation for developing HCMV subunit vaccines.关键词
人巨细胞病毒(HCMV)/包膜糖蛋白B(gB)/中和表位/单克隆抗体(mAb)/酶联免疫吸附反应(ELISA)/免疫反应性Key words
Human Cytomegalovirus (HCMV)/envelope glycoprotein B (gB)/neutralizing epitope/monoclonal antibody (mAb)/enzyme-linked immuno sorbent assay (ELISA)/immunoreactivity分类
生物科学引用本文复制引用
何赛,郭向荣,刘如石,廖旻晶,靳晓瑞,邱义兰,李晓丹,郑姣,唐娜,罗焕,梁湘辉..人巨细胞病毒包膜糖蛋白B主要中和表位单克隆抗体的制备[J].生命科学研究,2017,21(1):16-22,7.基金项目
湖南省科技计划重点研发计划(2015SK20332,2011SK2014) （2015SK20332,2011SK2014）
湖南省教育厅高等学校科研项目(15C0829) （15C0829）
湖南省自然科学基金项目(09JJ5027) （09JJ5027）
