中国兽医科学2025，Vol.55Issue(9)：1174-1181,8.DOI:10.16656/j.issn.1673-4696.2025.0174
猴痘病毒gp085蛋白生物信息学分析及其多克隆抗体制备
Bioinformatics analysis and polyclonal antibody preparation of monkeypox virus gp085 protein
摘要
Abstract
This study aims to predict and analyze the physicochemical properties,structure,and antigenic epitopes of the MPXV gp085 protein using bioinformatics analysis.Express and purify the gp085 recombinant protein,and prepare and analyze rabbits-derived polyclonal antibodies.Taking the MPXV gp085 gene as the research object,apply online websites to analyze its bioinformatics,use the prokaryotic expression system to express the gp085 recombinant protein,and the purification efficacy was confirmed by SDS-PAGE after purification.The purified gp085 protein was used to immunize rabbits for the production of polyclonal antibodies.The antibody titer was determined by indirect ELISA,and the specificity of the polyclonal antibodies was evaluated by Western-blot.Bioinformatics analysis showed that the MPXV gp085 protein is a stable hydrophilic protein without transmembrane regions and signal peptides,with three linear epitopes.The gp085 homologous gene sequences of nine orthopoxviruses have high consistency,and MPXV has the closest genetic relationship with cowpox virus.The recombinant gp085 protein was mainly expressed as inclusion bodies,with an approximate size of 17 kDa.The results of indirect ELISA detection showed that the titer of polyclonal antibody reached 1∶51 200.The West-ern-blot results indicated that the polyclonal antibody could specifically recognize the gp085 pro-tein.The recombinant MPXV gp085 protein has good antigenicity and can stimulate the production of high titer serum antibody in immunized rabbits.This study provides biological materials and theoretical basis for further studying the biological functions of MPXV gp085 protein and the development of rapid diagnostic technology.关键词
猴痘病毒/gp085蛋白/生物信息学分析/原核表达/多克隆抗体Key words
monkeypox virus/gp085 protein/bioinformatics analysis/prokaryotic expression/poly-clonal antibody分类
农业科技引用本文复制引用
张亚岭,曾巧英,王振蕊,景伟,苏洋,陈国华,何小兵,房永祥,李小明,景志忠..猴痘病毒gp085蛋白生物信息学分析及其多克隆抗体制备[J].中国兽医科学,2025,55(9):1174-1181,8.基金项目
国家自然科学基金面上项目(32473069) （32473069）
"十四五"国家重点研发计划项目(2023YFD1802501) （2023YFD1802501）
兰州市科技计划项目(2023-1-38) （2023-1-38）
甘肃省科技重大专项(23ZDNA007) （23ZDNA007）
国家动物疫情监测与防治专项(125161031) （125161031）
甘肃省青年科技基金项目(22JR5RA780) （22JR5RA780）
