液相ELISA法测定人血浆抗英夫利西单抗抗体及在克罗恩病患者中应用  

Measurements of antibodies to infliximab in plasma by solution ELISA and its application in patients of Crohn's disease

作　　者：朱扣柱 丁肖梁[1] 王燕 陈卫昌[3] 缪丽燕[1] ZHU Kouzhu;DING Xiaoliang;WANG Yan;CHENG Weichang;MIAO Liyan(Department of Pharmacy,First Affiliated Hospital,Soochow University,Jiangsu Suzhou 215006,China;Department of pharmacy,Affiliated Children's Hospital,Jiangnan University(Wuxi Children's Hospital),Jiangsu Wuxi 214023,China;Department of Gastroenterology,First Affiliated Hospital,Soochow University,Jiangsu Suzhou 215006,China)

机构地区：[1]苏州大学附属第一医院药学部,江苏苏州215006 [2]江南大学附属儿童医院(无锡市儿童医院)药学部,江苏无锡214023 [3]苏州大学附属第一医院消化内科,江苏苏州215006

出　　处：《中国医院药学杂志》2024年第15期1744-1749,共6页Chinese Journal of Hospital Pharmacy

基　　金：苏州市姑苏人才项目(编号:GSWS2019001);国家自然科学基金青年项目资助(编号:82003857)。

摘　　要：目的:建立人血浆抗英夫利西单抗抗体(antibodies to infliximab, ATI)免疫分析方法并进行方法学验证。方法:用甘氨酸酸解英夫利西单抗(infliximab, IFX)与ATI结合形成的复合物,然后用生物素标记IFX和辣根过氧化物酶(HRP)标记IFX结合ATI形成桥联复合物,再将溶液转移到链霉亲和素包被的孔板上进行检测。方法建立后采用51例未曾使用过IFX的克罗恩病患者血浆进行验证。结果:检测ATI的ELISA方法经验证筛选临界值为1.16,确证临界值为17.21%,滴度临界值为1.34,批内和批间精密度均小于20%,灵敏度达50 ng·mL^(-1),当阳性对照抗体质量浓度为100 ng·mL^(-1)时,药物耐受水平为12.5μg·mL^(-1)。结论:成功建立并验证了检测人血浆ATI的免疫分析方法,适合临床开展治疗药物检测。OBJECTIVE To develop and validate a solution enzyme-linked immunosorbent assay(ELISA)method for detecting antibodies to infliximab(ATI).METHODS The infliximab(IFX)/ATI complexes were dissociated by glycine acid,then biotin-IFX and HRP-IFX conjugated with ATI and then resulting bridging complexes were transferred to a streptavidin matrix coated 96-well plate.After an establishment of ATI assay,plasma samples from 51 IFX-naive Crohn's disease(CD)patients were utilized for method validation.RESULTS Based upon a panel of plasma samples from 51 IFX-naive CD patients,the screening cut-point was 1.16(S/N).The confirmatory cut-point was determined as a decrease in signal of>17.21%.The titer cut-point was 1.34.Both intra-and inter-assay precisions were<20% of CV.The relative sensitivity of this assay was 50ng·mL^(-1).Drug-tolerance at the concentration of 100 ng·mL^(-1) positive control was≤12.50μg·mL^(-1) IFX.The relative sensitivity of this assay was 50 ng·mL^(-1).CONCLUSION The immunoassay for measuring ATI has been successfully developed and validated,this method can be used for therapeutic drug monitoring.

关 键 词：英夫利西单抗 抗英夫利西单抗抗体 克罗恩病 酶联免疫吸附分析法 

分 类 号：R969.1[医药卫生—药理学]