色谱法
乌斯特基努马
分析物
质谱法
串联质谱法
化学
检出限
液相色谱-质谱法
治疗药物监测
免疫分析
抗体
医学
药理学
肿瘤坏死因子α
药代动力学
内科学
免疫学
阿达木单抗
作者
Nina Scheffe,Rupert Schreiner,A Thomann,Peter Findeisen
出处
期刊:Therapeutic Drug Monitoring
[Ovid Technologies (Wolters Kluwer)]
日期:2020-08-01
卷期号:42 (4): 572-577
被引量:2
标识
DOI:10.1097/ftd.0000000000000734
摘要
Background: Ustekinumab (UST) is a human monoclonal antibody used to treat moderate-to-severe Crohn disease by blocking the interleukin-12/23 pathway. Although an optimized therapeutic concentration of UST is associated with clinical response and improved prognosis, the availability of clinical laboratory methods for UST monitoring is limited. Furthermore, the commercially available methods are immunoassays that are prone to interference of antidrug antibodies. This study aimed to develop a liquid chromatography-tandem mass spectrometry method for quantification of UST in human serum specimens. Methods: A tryptic peptide that is specific to the heavy chain variable region of UST was selected. Quantification of UST was performed by selective reaction monitoring on a quadrupole TQ-XS with an internal standard. After digestion with trypsin, peptides were separated by reverse-phase C18 liquid chromatography; peptides were detected by MS/MS, and analyte to internal standard peak area ratios were used for the quantification. Finally, serum samples from patients treated with UST were collected at trough levels (n = 66). Results: The assay showed a broad dynamic range with linearity between 0.4 and 20 mg/L (R 2 = 0.995). The lower limit of quantification was found to be 0.4 mg/L. The reproducibility was tested with 3 different UST concentrations (2, 8, and 16 mg/L). The coefficients of intra-assay and interassay variations were 2.2%–4.0% and 2.7%–5.3%, respectively. UST serum concentrations of 2–16 mg/L were stable for up to 14 days when specimens were left at room temperature (20°C). Conclusions: The newly developed LC/MS-based method was shown to be feasible for UST quantification. This analytical approach may lead to individualized dosing and improved patient care.
科研通智能强力驱动
Strongly Powered by AbleSci AI