猪IL-1β单克隆抗体制备及其抗炎症反应活性  

Preparation and Anti-inflammatory Activity of Porcine IL-1βMonoclonal Antibody

作　　者：李璠 孙海凤[1] 孙萌 高雁怩 孙杨杨[1] 张路捷 白娟[1] 姜平[1] LI Fan;SUN Haifeng;SUN Meng;GAO Yanxiao;SUN Yangyang;ZHANG Lujie;BAI Juan;JIANG Ping(Key Laboratory of Animal Bacteriology of Ministry of Agriculture and Rural Affairs,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学/农业农村部动物细菌学重点实验室,南京210095

出　　处：《畜牧兽医学报》2025年第2期890-899,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金项目(32230103);国家生猪产业技术体系专项(CARS-35);江苏省自主创新研发基金项目(CX(22)1011)。

摘　　要：IL-1β可介导炎症病理,阻断IL-1活性具有重要抗炎治疗作用。本研究通过构建猪IL-1β原核表达质粒,进行大肠杆菌表达和蛋白纯化,制备重组蛋白。采用细胞杂交瘤技术制备获得4株稳定分泌IL-1β单克隆抗体杂交瘤细胞株(1E2、2H3、5H3及7E7);4株单抗腹水ELISA效价高达1∶1024000;5H3单抗识别的抗原表位为149 DLKREVVFCM 158;1E2、2H3和7E7单抗所识别的抗原表位为202 KRYPKRD 208。脂多糖(LPS)小鼠炎症模型抗炎试验结果显示,相对LPS对照组,IL-1β单抗处理组小鼠血清NO、TNF-α、IL-6和IL-1β含量及临床症状评分显著降低,表明4株单抗均有较好抗炎效果,其中1E2单抗抗炎作用最优,为猪IL-1β阻断药物研究奠定了重要基础。IL-1βmediates inflammatory pathology,and blocking IL-1 activity has important anti-inflammatory effects.In this study,recombinant protein was prepared by constructing porcine IL-1βprokaryotic expression plasmid,expression by Escherichia coli and protein purification.Four hybridoma cell lines(1E2,2H3,5H3 and 7E7)with stable IL-1βmonoclonal antibody secretion were prepared by cell hybridoma technique.The ELISA titer of the 4 mAbs in ascites were up to 1:1024000.The epitope of 5H3 mAb was 149 DLKREVVFCM 158.The epitope recognized by 1E2,2H3 and 7E7 mAb was 202 KRYPKRD 208.The anti-inflammatory test results of lipopolysaccharide(LPS)mice inflammation model showed that compared with LPS control group,serum NO,TNF-α,IL-6 and IL-1βcontents and clinical symptom score of mice treated with IL-1βmonoclonal antibody were significantly reduced,indicating that all the four strains of monoclonal antibody had better anti-inflammatory effect,and 1E2 monoclonal antibody had the best anti-inflammatory effect.It laid an important foundation for the study of porcine IL-1βblocking drugs.

关 键 词：IL-1Β 单克隆抗体 炎症治疗 

分 类 号：S852.659.6[农业科学—基础兽医学]