Antibody-free quantification of serum infliximab using LC-MS/MS

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-08-14 DOI:10.1016/j.jpba.2025.117117

Xiaoshuang He , Juntao Zhao , Xiaoli Ma , Wenkai Chen , Shu Chen , Kaifeng Shi , Jian Wei , Zhichao Gu , Jie Fang , Fengmei Hu , Xiaolan Bian , Jing Sun , Yuan Xiao , Qiuya Lu

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引用次数: 0

Abstract

Background

Infliximab (IFX) is the first biologic to be approved to treat inflammatory bowel disease (IBD). Serum IFX concentrations can be correlated with the clinical prognosis of IBD. Liquid chromatography–tandem mass spectrometry (LC-MS/MS) is an alternative to clinical immunoassays owing to its higher selectivity and sensitivity. An efficient antibody-free LC-MS/MS method was developed in this study to determine serum IFX levels.

Methods

Protein G–coated magnetic beads were used to capture IFX and the internal standard (cadonilimab [CADO]) in human serum. After enrichment, IFX and CADO were digested using trypsin, and signature peptides were selected for subsequent LC-MS/MS analysis. The results were compared with those from the corresponding immunoassay.

Results

The selected peptides had good specificity, and the total precision was within 10.5 %. The accuracy was between 92.8 % and 97.6 % in the linear range of 0.5–100 mg/L. The results from the sample assay were in agreement with those from the immunoassay.

Conclusion

Determination of IFX using LC-MS/MS is more sensitive and robust than other analytical methods. Therefore, the method reported in this study shows potential for use in therapeutical drug monitoring and developing individualized treatment modalities.

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LC-MS/MS法测定英夫利昔单抗血清的无抗体含量

dinfliximab （IFX）是首个被批准用于治疗炎症性肠病（IBD）的生物制剂。血清IFX浓度与IBD的临床预后相关。液相色谱-串联质谱法（LC-MS/MS）具有较高的选择性和灵敏度，是临床免疫测定的一种替代方法。本研究建立了一种高效的无抗体LC-MS/MS检测血清IFX水平的方法。方法采用蛋白g包被磁珠捕获人血清中的IFX和内标卡多尼单抗。富集后，用胰蛋白酶消化IFX和CADO，并选择特征肽进行LC-MS/MS分析。将结果与相应的免疫分析结果进行比较。结果所选肽具有良好的特异性，总精密度在10.0 %以内。在0.5 ~ 100 mg/L的线性范围内，准确度为92.8 % ~ 97.6% %。样品分析的结果与免疫分析的结果一致。结论LC-MS/MS法测定IFX具有较高的灵敏度和鲁棒性。因此，本研究中报告的方法显示了在治疗药物监测和开发个体化治疗模式方面的潜力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of pharmaceutical and biomedical analysis 医学-分析化学

CiteScore

6.70

自引率

5.90%

发文量

588

审稿时长

37 days

期刊介绍： This journal is an international medium directed towards the needs of academic, clinical, government and industrial analysis by publishing original research reports and critical reviews on pharmaceutical and biomedical analysis. It covers the interdisciplinary aspects of analysis in the pharmaceutical, biomedical and clinical sciences, including developments in analytical methodology, instrumentation, computation and interpretation. Submissions on novel applications focusing on drug purity and stability studies, pharmacokinetics, therapeutic monitoring, metabolic profiling; drug-related aspects of analytical biochemistry and forensic toxicology; quality assurance in the pharmaceutical industry are also welcome. Studies from areas of well established and poorly selective methods, such as UV-VIS spectrophotometry (including derivative and multi-wavelength measurements), basic electroanalytical (potentiometric, polarographic and voltammetric) methods, fluorimetry, flow-injection analysis, etc. are accepted for publication in exceptional cases only, if a unique and substantial advantage over presently known systems is demonstrated. The same applies to the assay of simple drug formulations by any kind of methods and the determination of drugs in biological samples based merely on spiked samples. Drug purity/stability studies should contain information on the structure elucidation of the impurities/degradants.