抗重组人血清白蛋白抗体的检测方法建立及验证  

Establishment and verification of a method for detection of anti-drug antibody against recombinant human serum albumin

作　　者：杨淑涵 刘海涌 贾向阳 刘丽[1] 王平 李曙芳[4] 潘东升[1] 李波[1] YANG Shu-han;LIU Hai-yong;JIA Xiang-yang;LIU Li;WANG Ping;LI Shu-fang;PAN Dong-sheng;LI Bo(Beijing Key Laboratory of Non-clinical Safety Evaluation of Drugs,National Center for Safety Evaluation of Drugs,National Institutes for Food and Drug Control,Bejing 100176,China;Bejing Xinajin Biotechnology Company,Bejing 100176,China;Shenzhen Protgen Ltd.,Shenzhen 518048,China;Shanxi Key Laboratory of Drug Toxicology and Drug for Radiation Injury,China Institute for Radiation Protection Research,Taiyuan 030006,China)

机构地区：[1]中国食品药品检定研究院国家药物安全评价监测中心,药物非临床安全评价研究北京市重点实验室,北京100176 [2]北京新艾进生物科技有限公司,北京100176 [3]深圳普罗吉医药科技有限公司,深圳518048 [4]药物毒理与放射损伤药物山西省重点实验室,中国辐射防护研究院,太原030006

出　　处：《中国新药杂志》2023年第18期1866-1873,共8页Chinese Journal of New Drugs

基　　金：中国辐射防护研究院自主科研项目平台开放基金(CIRP-DTRI20220201)。

摘　　要：目的:药物耐受以及内源性交叉反应是影响抗药抗体检测的主要因素,因此建立能够耐受较高药物浓度且可避免内源性交叉反应的检测方法是抗药抗体分析中需要解决的主要问题。本研究目的是建立抗重组人血清白蛋白抗体的检测方法并进行验证。方法:通过酸化及纳米磁珠富集提取抗重组人血清白蛋白抗体,然后采用桥连法进行抗体检测,最后对建立的方法进行验证。结果:经过该方法处理后,抗体回收率>80%,方法灵敏度可达到0.5μg·mL^(-1),批间精密度和批内精密度均<20%,该方法最高可耐受2 mg·mL^(-1)的药物。结论:本研究建立了抗重组人血清白蛋白抗体检测方法,该方法抗体回收率高,具有良好的灵敏度和精密度,可达到mg级耐药,且可有效去除内源性物质干扰,既可用于抗药抗体检测,也可用于抗体药物提取,进而用于药动学分析。Objective:Drug resistance and endogenous cross-reactivity are the main factors affecting the detection of anti-drug antibody(ADA),so the establishment of method that can tolerate higher drug concentration and avoid endogenous cross-reactivity is a major problem to be solved in immunogenicity analysis.The aim of this study was to establish and verify a method for detection of ADA against recombinant human serum albumin(rHSA).Methods:Anti-rHSA antibody was extracted by acidification and enrichment with magnetic nanobeads and then detected using a bridging method.Finally,the established method was validated.Results:The recovery of the antibody was greater than 80%by using the established method.The sensitivity of the method could reach 0.5μg·mL^(-1)and the intra-and inter-batch RSDs were less than 20%.In addition,the method could resist drug of up to 2 mg·mL^(-1).Conclusion:This study established a method for the detection of anti-rHSA antibody.The method has high antibody recovery and good performances of sensitivity and precision.It can also achieve mg-level drug-resistance and effectively remove the interference of endogenous substances,which can be applied for the detection of ADA,extraction of antibody drugs and further analysis of drug metabolism.

关 键 词：抗药抗体 耐药性 交叉反应 血清白蛋白 富集提取 

分 类 号：R927.1[医药卫生—药学]