病毒感染标志物MxA抗体的制备及基于计算机模拟的亲和力提升  

Preparation of MxA Antibodies as Biomarkers for Viral Infections and Affinity Enhancement via Computational Simulation

作　　者：衡玉 党英萍 王競 徐小霞 潘能科 胡伟 唐娜 宋其涛 易维京 谭兵 HENG Yu;DANG Yingping;WANG Jing;XU Xiaoxia;PAN Nengke;HU Wei;TANG Na;SONG Qitao;YI Weijing;TAN Bing(Zybio Inc.,Chongqing 400080,China;Chongqing Essence Biological Engineering Co.,Ltd.,Chongqing 400080,China;Chongqing Center for Drug Evaluation and Certification,Chongqing 400000,China)

机构地区：[1]中元汇吉生物技术股份有限公司,重庆400080 [2]重庆艾生斯生物工程有限公司,重庆400080 [3]重庆市药品技术审评查验中心,重庆400000

出　　处：《生物化工》2025年第1期38-42,共5页Biological Chemical Engineering

基　　金：重庆市自然科学基金面上项目(CSTB2023NSCQ-MSX0368);重庆市技术创新与应用发展专项重大项目(CSTB2023TIAD-STX0011);重庆市技术创新与应用发展专项重点项目(CSTB2021TIAD-KPX0044)。

摘　　要：目的:开发针对粘液病毒抗性蛋白A(Myxovirus resistance protein A,MxA)的高亲和力抗体,旨在满足MxA体外检测试剂开发对抗体原料的高灵敏度需求。方法:通过传统杂交瘤抗体开发技术制备抗MxA抗体,结合计算机模拟手段,运用虚拟氨基酸突变等方法,对抗体进行结构优化。结果:计算机模拟技术确定抗原-抗体关键氨基酸位点为重链第37、38、57、66、106、108、117位,轻链第34、36、38、56位。对这些位点进行虚拟氨基酸突变,得到轻链ASN36→ARG突变抗体。亲和力分析显示该突变抗体比野生型抗体亲和力提高2个数量级,KD值达到5.93×10^(-11) mol/L。结论:本文成功开发了抗MxA单克隆抗体,并通过计算机模拟技术使该抗体亲和力提升了2个数量级,为抗体的体外亲和力成熟提供了理论指导。Objective:Develop high-affinity antibodies against myxovirus resistance protein A(MxA)to meet the high sensitivity requirements of antibody raw materials for the development of MxA in vitro detection reagents.Methods:Anti-MxA antibodies are prepared by traditional techniques,combined with computational modeling,antibodies with theoretically inferred high-affinity structure are established and evaluated.Results:Computer simulation techniques determined that the key amino acid sites of the antigen-antibody are amino acids at positions 37,38,57,66,106,108,and 117 of the heavy chain,and amino acids at positions 34,36,38,and 56 of the light chain.Virtual amino acid mutations are carried out on these sites,and an antibody with the ASN36 to ARG mutation in the light chain is obtained.Analysis show that the affinity of this mutant antibody is improved by two orders of magnitude compared with that of the wild-type antibody,with a KD value reaching 5.93×10^(-11) mol/L.Conclusion:An anti-MxA monoclonal antibody is successfully developed,and the affinity of this antibody is enhanced by two orders of magnitude through computer simulation techniques,providing theoretical guidance for the in vitro affinity maturation of antibodies.

关 键 词：MXA 抗体 计算机模拟 亲和力提升 体外诊断 

分 类 号：R446.61[医药卫生—诊断学]