抗平颏海蛇短链神经毒素全人源单克隆抗体的筛

发布时间：2018-05-27 01:06

本文选题：平颏海蛇 + 短链神经毒素　；参考：《解放军医学杂志》2017年07期

【摘要】：目的重组表达3种平颏海蛇短链神经毒素,利用噬菌体抗体库筛选制备全人源抗毒素单克隆抗体并评估其生物活性。方法以重组表达纯化的神经毒素蛋白作为抗原,从自主构建的噬菌体抗体库中筛选获得噬菌体抗体,经序列测定确定其抗体类型后构建完整抗体。利用真核表达系统表达纯化抗体后鉴定其抗原结合活性、生化特性及药代动力学特性。在体内验证抗体药物的抗毒素作用。结果经过4轮筛选,获得2株能同时结合3种神经毒素的噬菌体单链抗体,并将其制备成完整的抗体,进一步证实所获得的抗体可特异性结合3种抗原。2株全人源抗体均可拮抗神经毒素对昆明小鼠的致死作用。结论利用重组神经毒素和噬菌体抗体库技术成功获得了特异性抗平颏海蛇短链神经毒素的全人源治疗性抗体。
[Abstract]:Objective to express three kinds of short chain neurotoxins of sea snake, and to prepare monoclonal antibodies against human antitoxin by phage antibody library and evaluate their biological activity. Methods the phage antibody was obtained from the self-constructed phage antibody library using recombinant and purified neurotoxin protein as antigen. The antibody type was determined by sequencing and the complete antibody was constructed. The antigen-binding activity, biochemical characteristics and pharmacokinetics of the purified antibody were identified by eukaryotic expression system. To test the antitoxin effect of antibody drugs in vivo. Results after four rounds of screening, two phage scFv, which could bind to 3 neurotoxins simultaneously, were obtained and prepared into complete antibodies. It was further confirmed that the obtained antibodies could specifically bind to all human antibodies of 3 antigens and could antagonize the lethal effect of neurotoxin on Kunming mice. Conclusion the recombinant neurotoxin and phage antibody library were used to obtain the specific human therapeutic antibody against the short chain neurotoxin of sea snake.
【作者单位】：第二军医大学基础医学部生物物理学教研室;
【基金】：国家自然科学基金(81602690)~~
【分类号】：R392

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