摘要
目的建立测定人血清中阿达木单抗药物浓度的酶联免疫分析(ELISA)方法,测定强直性脊柱炎(AS)患者体内阿达木单抗血清稳态谷浓度并研究个体间和个体内的变异。方法固相微孔板上包被人肿瘤坏死因子α(hTNF-α)作为捕获抗原,加入待检测样品,以羊抗人IgG-HRP为检测抗体,TMB为底物进行显色并测定450 nm处的吸光度。47例经标准剂量阿达木单抗注射液治疗的AS患者,采集治疗后12周至24周稳态谷浓度血样,测定药物浓度。结果该方法定量范围为0. 63~20. 00μg·mL^-1,批间和批内精密度均不超过15%,平均回收率<110%;47例患者治疗第12周时血清中位谷浓度为10. 15μg·mL^-1(IQR:4. 36~13. 31μg·mL^-1),浓度范围为0. 00~23. 71μg·mL^-1,个体间变异为63%;43例患者中个体内变异中位数为11%(IQR:8~17μg·mL^-1)。结论本研究建立的方法稳定性好,适用于临床样本的检测。阿达木单抗治疗稳态谷浓度个体间变异较大而个体内变异较小,基于稳态谷浓度的"被动治疗药物监测"将有助于给药方案调整。
OBJECTIVE To establish an enzyme-linked immunosorbent assay( ELISA) method for determining the concentration of adalimumab in human serum to determine the steady-state trough concentration of adalimumab in patients with ankylosing spondylitis( AS) and to study inter-and intraindividual variation. METHODS Human tumor necrosis factor α( hTNF-α) was coated on the solid phase microplate as the capture antigen,and the sample to be tested was added. The goat anti-human IgG-HRP was used as the detection antibody and TMB was used for color development. The absorbance at 450 nm was finally determined. Forty-seven patients with AS treated with standard dose of adalimumab injection were included. Blood samples of steady-state after 12 to 24 weeks’ treatment were collected to determine trough drug concentrations. RESULTS The quantitative range of the method was 0. 63-20. 00 μg·mL^-1. The inter-assay and intra-assay precisions were not more than 15 % and the average recovery was < 110 %. The median serum concentration of 47 patients was 10. 15 μg · mL^-1( IQR: 4. 36 ~ 13. 31μg·mL^-1) and ranged from 0. 00-23. 71 μg·mL^-1 with an inter-individual variation of 63 %;the median variation of intraindividuals within 43 patients was 11 %( IQR: 8 %-17 %). CONCLUSION This method is stable and is suitable for the detection of clinical samples. Pronounced inter-but not intraindividual variation was observed in adalimumab trough levels in steady state. The " reaction therapeutic drug monitoring" based on the steady-state trough concentration will facilitate the adjustment of the dosing regimen.
作者
朱瑞芳
李香莲
丁肖梁
武剑
缪丽燕
ZHU Rui-fang;LI Xiang-lian;DING Xiao-liang;WU Jian;MIAO Li-yan(Department of Clinical Pharmacology Research Lab, The First Affiliated Hospital of Soochow University, Suzhou 215006, China;Department of Rheumatology, The First Affiliated Hospital of Soochow University, Suzhou 215006, China;College of Pharmaceutical Sciences, Soochow University, Suzhou 215100, China)
出处
《中国药学杂志》
CAS
CSCD
北大核心
2019年第13期1086-1091,共6页
Chinese Pharmaceutical Journal
基金
江苏省医学重点人才项目资助(ZDRCA2016048)
苏州市药物临床研究与个体化治疗重点实验室项目资助(SZS201719)
