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0.63~20.00 μg·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10.15 μg·mL-1(IQR:4.36~13.31 μg·mL-1),浓度范围为0.00~23.71 μg·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:8~17 μg·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。
Abstract
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:4.36~13.31 μg·mL-1) and ranged from 0.00-23.71 μg·mL-1 with an inter-individual variation of 63%; the median variation of intra-individuals 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.
关键词
阿达木单抗 /
免疫分析法 /
个体内和个体间变异
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Key words
adalimumab /
ELISA /
inter- and intraindividual variation
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中图分类号：
R969.1
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脚注
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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)
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