李坤
李坤，男，1986年生，博士，副研究员，中国农业科学院研究生院硕导，陇原青年英才、兰州兽医研究所“所级”培育青年英才。聚焦兽医免疫生物学，建立了家畜源B细胞抗体库制备技术平台，用于解析病毒抗原构效关系，揭示保护性免疫应答机制。创立口蹄疫病毒（FMDV）、猪繁殖与呼吸综合征（PRRSV）等病毒的抗体库；首次揭示FMDV不同血清型间存在保守抗原结构与病毒粒子结构弱点，证实免疫O、A型FMDV疫苗可以诱导针对Asia1的交叉中和抗体应答。基于单B细胞抗体创新FMDV、PRRSV等病毒的检测方法。主持国自然科学基金青年项目、面上项目、十四五国家重点研发计划子课题共3项；以第一/通讯作者在PLOS Pathogens、Journal of Virology等期刊发表论文9篇；授权国际/国家发明专利11项。 一、研究方向：围绕抗原和抗体分子设计开展以下三方面研究。 1. B细胞抗体应答和抗体库解析-构效关系 围绕FMDV等烈性传染性病原，建立B细胞抗体库，开展抗原构效关系研究。聚焦抗体克隆型的定义，结合氨基酸大语言模型，开展抗体的分类研究，解析“抗原表位-抗体序列”之间映射关系，绘制病毒抗原结构图谱。 2. 结构抗原分子设计-结构疫苗 聚焦和优化设计载体蛋白，聚焦保守抗原结构，抽提关键抗原表位，使用扩散生成模型产生全新抗原分子。采用Alphafold3等结构模拟软件对新抗原分子进行结构吻合性评价，生产性能指标评价，鉴定候选分子。体外表达候选分子，ELISA评价抗原分子与广谱中和抗体结合能力。构建不同结构特征的载体模型，加载优势表位抗原，形成结构疫苗分子。 3. 反向抗体生成与优化设计 基于Transformer架构，结合蛋白质大预言生成模型（RFantibody、IgGM和Germinal等），指定位点关键残基，从头生成新的抗体分子。通过体外合成技术，构建抗体展示文库，筛选获得功能性靶标抗体。 二、发表文章 1.Huang S, Wu S, Li F, Li P, Sun P, Cao Y, Bao H, Dong K, Yang J, Zhang H, Zhao Q, Sun Y, Li D, Bai X, Fu Y, Yuan H, Ma X, Zhao Z, Zhang J, Wang J, Liu Z, Peng Y, Li K, Hua J, Lu Z, Lei D, Zhang Q. Porcine B cell receptor repertoire uncovers balanced recognition of antigenic structures on serotype Asia1 foot-and-mouth disease virus. Plos Pathogens, 2026. 已接受. 2.Huang S, Yang J, Li F, Zhang H, Cao Y, Zha J, Bao H, Li P, Liang L, Liu Z, Li K, Lu Z, Hua J, Zhang Q. Two residue mutations in VP1 of FMDV serotype A cause significant antigenic variation via reorientation of G-H loop revealed by lineage-specific neutralizing antibodies from the natural hosts. Vet Microbiol. 2025 Dec;311:110748. 3.Sun Y, Huang S, Li F, Huang S, Li P, Zhao Q, Wang T, Bao H, Fu Y, Sun P, Bai X, Yuan H, Ma X, Zhao Z, Zhang J, Wang J, Li D, Zhang Q, Cao Y, Li K, Lu Z, Fan H. Porcine antibodies reveal novel non-neutralizing universal epitopes on FMDV and their overlaps with neutralization sites. Vet Microbiol. 2025 Apr;303:110440. 4.Cao Y, Li F, Xing X, Zhang H, Zhao Q, Sun P, Fu Y, Li P, Ma X, Zhang J, Zhao Z, Yuan H, Wang J, Wang T, Bao H, Bai X, Li D, Zhang Q, Li K*, Lu Z*. Preparation and application of porcine broadly neutralizing monoclonal antibodies in an immunoassay for efficiently detecting neutralizing antibodies against foot-and-mouth disease virus serotype O. Microbiol Spectr. 2025 Feb 4;13(2):e0223424. 5.Zhao Q, Li F, Huang S, Xing X, Sun Y, Li P, Bao H, Fu Y, Sun P, Bai X, Yuan H, Ma X, Zhao Z, Zhang J, Wang J, Wang T, Li D, Zhang Q, Qian P, Cao Y*, Li K*, Li X*, Lu Z*. Remapping the spatial distribution of neutralizing sites and their immunodominance on the capsid of different topotypes of FMDV serotype O by site-directed competitive ELISA for detection of neutralizing antibodies. Microbiol Spectr. 2025 Jun 3;13(6): e0334424. 6.Li F, Wu S, Lv L, Huang S, Zhang Z, Zerang Z, Li P, Cao Y, Bao H, Sun P, Bai X, He Y, Fu Y, Yuan H, Ma X, Zhao Z, Zhang J, Wang J, Wang T, Li D, Zhang Q, He J, Liu Z, Lu Z, Lei D, Li K*. Discovery, recognized antigenic structures, and evolution of cross-serotype broadly neutralizing antibodies from porcine B-cell repertoires against foot-and-mouth disease virus. PLoS Pathog. 2024, 20(10): e1012623. 7.王莉, 李坤, 黄书伦, 李凤娟, 王省, 卢曾军, 刘在新. 口蹄疫病毒O型东南亚拓扑型抗原变异研究. 中国农业科学, 2024, 57(15): 3093-3104 8．Li K#, He Y#, Wang L, Li P, Huang S, Zhou S, Zhu G, Song Y, Li Y, Wang S, Zhang Q, Sun P, Bao H, Bai X, Zhao Z, Cao Y*, Lou Z*, Lu Z*, Liu Z. Conserved antigen structures and antibody-driven variations on foot-and-mouth disease virus serotype A revealed by bovine neutralizing monoclonal antibodies. Plos Pathogens, 2023,19(11), e1011811. 9. Cao Y, Li K, Xing X, Zhu G, Fu Y, Bao H, Bai X, Sun P, Li P, Zhang J, Ma X, Wang J, Zhao Z, Li D, Liu Z, Lu Z. Development and Validation of a Competitive ELISA Based on Bovine Monoclonal Antibodies for the Detection of Neutralizing Antibodies against Foot-and-Mouth Disease Virus Serotype A. J Clin Microbiol. 2022, 60(4): e0214221. 10. He Y#, Li K#, Wang L, Sun Z, Cao Y, Li P, Sun P, Bao H, Zhou S, Wang S, Bai X, Liu X, Zhao L, Fan X, Liu Z, Lu Z, Yang C, Lou Z. Structures of Foot-and-Mouth Disease Virus with Bovine Neutralizing Antibodies Reveal the Determinant of Intraserotype Cross-Neutralization. J Virol. 2021 Nov 23;95(24):e0130821. 11. Li K#, He Y#, Wang L, Li P, Wang S, Sun P, Bao H, Cao Y, Liu X, Zhu G, Song Y, Bai X, Ma X, Fu Y, Yuan H, Zhang J, Wang J, Chen Y, Li D, Lou Z*, Liu Z*, Lu Z*. Two Cross-Protective Antigen Sites on Foot-and-Mouth Disease Virus Serotype O Structurally Revealed by Broadly Neutralizing Antibodies from Cattle. J Virol. 2021 Oct 13;95(21):e0088121. 12．Li K, Zhu G, Zhou S, Sun P, Wang H, Bao H, Fu Y, Li P, Bai X, Ma X, Zhang J, Li D, Chen Y, Liu Z, Cao Y, Lu Z. Isolation and characterization of porcine monoclonal antibodies revealed two distinct serotype-independent epitopes on VP2 of foot-and-mouth disease virus. J Gen Virol. 2021 Jul;102(7). 13. Li K#, Bao H#, Wei G, Li D, Chen Y, Fu Y, Cao Y, Li P, Sun P, Bai X, Ma X, Zhang J, Lu Z, Liu Z. Molecular vaccine prepared by fusion of XCL1 to the multi-epitope protein of foot-and-mouth disease virus enhances the specific humoural immune response in cattle. Appl Microbiol Biotechnol. 2017 Nov;101(21):7889-7900. 三、联系方式 电话: 15393115826
