| CMG2-Fc 融合蛋白突变体筛选及中和炭疽毒素活性分析 |
| |
| 引用本文： | 刘宗伟,任雨春,郗永义,岳俊杰,张连成,邵勇,高丽华,胡显文,周艳荣,吴晓洁,陈红星.CMG2-Fc 融合蛋白突变体筛选及中和炭疽毒素活性分析[J].现代生物医学进展,2016,16(9):1601-1605. |
| |
| 作者姓名： | 刘宗伟 任雨春 郗永义 岳俊杰 张连成 邵勇 高丽华 胡显文 周艳荣 吴晓洁 陈红星 |
| |
| 作者单位： | 军事医学科学院生物工程研究所；常熟市公安局DNA 实验室；滕州市工人医 |
| |
| 基金项目： | 国家自然科学基金项目（81202445） |
| |
| 摘 要： | 目的:筛选能有效中和炭疽毒素和抵抗炭疽毒素损伤细胞的CMG2-Fc(炭疽毒素受体II-人免疫球蛋白Fc段融合蛋白)突变体。方法:运用FoldX等计算软件分析CMG2与PA晶体学结构,设计能提高CMG2-PA亲和力的突变体分子,并与人IgG1Fc片段构成融合基因,转染CHO-S细胞并通过亲和层析获得CMG2-Fc突变体蛋白,通过亲和力检测和细胞保护实验分析各突变体中和炭疽毒素能力。结果:筛选并表达了8个CMG2-Fc突变体分子,亲和力实验显示其中E117Q突变可明显提高CMG2-Fc与PA的亲和力(KD=1.35×10-11 mol/L),细胞保护实验提示E117Q突变能有效提高CMG2-Fc中和炭疽毒素能力(CMG2-Fc(E117Q)的IC50为15 ng/μL,而wt CMG2-Fc的IC50为50ng/μL)。结论:CMG2-Fc(E117Q)突变体分子可作为拮抗炭疽毒素损伤的炭疽治疗药物分子,进行进一步研究。
|
| 关 键 词： | 炭疽 炭疽毒素 CMG2 Fc融合蛋白 |
Design and Identify CMG2-Fc Mutants to Neutralizing Anthrax Toxin |
| |
| Abstract: | Objective:To identify CMG2-Fc mutant to neutralize anthrax toxin.Methods:CMG2-Fc Crystal structure was
analysed by FoldX software. CMG2-Fc mutants were designed and transfected in CHO-S cells, those mutants were purified by protein A
affinity chromatography. The abilities of CMG2-Fc mutants neutralizing anthrax toxin were analysed by affinity assay and toxin
neutralization assay in cells.Results:Eight CMG2-Fc mutants were designed and expressed, the affinity assay results show that E117Q
mutant could strengthen CMG2-PA binding, and cell protection assay results certify that CMG2-Fc (E117Q) mutant has superior ability
to neutralize toxin (IC50=15 ng/uL) than wtCMG2-Fc (IC50=50) ng/uL.Conclusion:CMG2-Fc (E117Q) was a superior anti-anthrax toxin
molecule and valuable to study more. |
| |
| Keywords: | Anthrax Anthrax toxin CMG2 Fc fusion protein |
| 本文献已被 CNKI 等数据库收录！ |
| 点击此处可从《现代生物医学进展》浏览原始摘要信息 |
| 点击此处可从《现代生物医学进展》下载免费的PDF全文 |
|
