| 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].,2016,16(9):1601-1605 |
| CMG2-Fc 融合蛋白突变体筛选及中和炭疽毒素活性分析 |
| Design and Identify CMG2-Fc Mutants to Neutralizing Anthrax Toxin |
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| DOI： |
| 中文关键词: 炭疽 炭疽毒素 CMG2 Fc融合蛋白 |
| 英文关键词: Anthrax Anthrax toxin CMG2 Fc fusion protein |
| 基金项目:国家自然科学基金项目（81202445） |
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| 中文摘要: |
| 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IgG1 Fc
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/uL，而wtCMG2-Fc 的IC50为50 ng/uL）。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。 |
| 英文摘要: |
| 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. |
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