Development of a time-resolved fluoroimmunoassay for rituximab and its application to therapeutic drug monitoring

IF 2.9 3区医学 Q2 MEDICAL LABORATORY TECHNOLOGY Clinica Chimica Acta Pub Date : 2025-02-15 Epub Date: 2025-01-20 DOI:10.1016/j.cca.2025.120142

Shangbin Kao , Xiaobin Liu , Juan Jin , Lin Zhang , Ting Shen , Jialong Wu , Yuan Qin , Xiumei Zhou , Xueqin Zhao , Liang Wang , Qiang He , Biao Huang

{"title":"Development of a time-resolved fluoroimmunoassay for rituximab and its application to therapeutic drug monitoring","authors":"Shangbin Kao , Xiaobin Liu , Juan Jin , Lin Zhang , Ting Shen , Jialong Wu , Yuan Qin , Xiumei Zhou , Xueqin Zhao , Liang Wang , Qiang He , Biao Huang","doi":"10.1016/j.cca.2025.120142","DOIUrl":null,"url":null,"abstract":"<div><h3>Background</h3><div>Rituximab pharmacokinetics in patients with membranous nephropathy (MN) exhibit significant interindividual variability. Accurate measurement of serum rituximab concentrations is essential for effective therapeutic monitoring. This study develops a highly sensitive time-resolved fluoroimmunoassay (TRFIA) for rituximab (rituximab-TRFIA) with a wide detection range, aimed at enhancing therapeutic drug monitoring in MN treatment.</div></div><div><h3>Methods</h3><div>A capture -type rituximab-TRFIA was developed using streptavidin-coated microplates, biotinylated anti-rituximab idiotypic antibodies, and Eu<sup>3+</sup>-labeled mouse anti-human IgG targeting the Fc fragment of rituximab. The assay was used to measure rituximab serum concentrations in MN patients treated with rituximab.</div></div><div><h3>Results</h3><div>The linear range of rituximab-TRFIA is 0.50 to 2500 ng/mL, with a limit of detection (LOD) of 0.062 ng/mL. The intra-assay coefficient of variation (CV) ranges from 1.97 % to 9.50 %, while the inter-assay CV ranges from 7.44 % to 9.99 %. The recovery rate ranges from 99.14 % to 107.75 %. No cross-reactivity was observed with other monoclonal antibody drugs (mAbs). The detection range is two orders of magnitude higher, and the sensitivity is six times greater than that of the enzyme-linked immunosorbent assay (ELISA). Rituximab-TRFIA showed strong consistency within the same measurement range compared to ELISA (P < 0.0001). The serum rituximab levels in MN patients were significantly higher than those in the control group (P < 0.0001). Among patients who did not achieve remission, 60 % to 70 % had insufficient drug concentrations, and rituximab pharmacokinetics followed the expected trends.</div></div><div><h3>Conclusions</h3><div>The rituximab-TRFIA provides high sensitivity, a wide detection range, and reliable performance for monitoring serum rituximab concentrations in MN patients, supporting personalized treatment strategies.</div></div>","PeriodicalId":10205,"journal":{"name":"Clinica Chimica Acta","volume":"568 ","pages":"Article 120142"},"PeriodicalIF":2.9000,"publicationDate":"2025-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Clinica Chimica Acta","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S000989812500021X","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/1/20 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"MEDICAL LABORATORY TECHNOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Background

Rituximab pharmacokinetics in patients with membranous nephropathy (MN) exhibit significant interindividual variability. Accurate measurement of serum rituximab concentrations is essential for effective therapeutic monitoring. This study develops a highly sensitive time-resolved fluoroimmunoassay (TRFIA) for rituximab (rituximab-TRFIA) with a wide detection range, aimed at enhancing therapeutic drug monitoring in MN treatment.

Methods

A capture -type rituximab-TRFIA was developed using streptavidin-coated microplates, biotinylated anti-rituximab idiotypic antibodies, and Eu³⁺-labeled mouse anti-human IgG targeting the Fc fragment of rituximab. The assay was used to measure rituximab serum concentrations in MN patients treated with rituximab.

Results

The linear range of rituximab-TRFIA is 0.50 to 2500 ng/mL, with a limit of detection (LOD) of 0.062 ng/mL. The intra-assay coefficient of variation (CV) ranges from 1.97 % to 9.50 %, while the inter-assay CV ranges from 7.44 % to 9.99 %. The recovery rate ranges from 99.14 % to 107.75 %. No cross-reactivity was observed with other monoclonal antibody drugs (mAbs). The detection range is two orders of magnitude higher, and the sensitivity is six times greater than that of the enzyme-linked immunosorbent assay (ELISA). Rituximab-TRFIA showed strong consistency within the same measurement range compared to ELISA (P < 0.0001). The serum rituximab levels in MN patients were significantly higher than those in the control group (P < 0.0001). Among patients who did not achieve remission, 60 % to 70 % had insufficient drug concentrations, and rituximab pharmacokinetics followed the expected trends.

Conclusions

The rituximab-TRFIA provides high sensitivity, a wide detection range, and reliable performance for monitoring serum rituximab concentrations in MN patients, supporting personalized treatment strategies.

查看原文

微信好友朋友圈 QQ好友复制链接

本刊更多论文

利妥昔单抗时间分辨荧光免疫测定法的建立及其在治疗药物监测中的应用。

背景：利妥昔单抗在膜性肾病（MN）患者中的药代动力学表现出显著的个体差异。准确测量血清利妥昔单抗浓度对有效的治疗监测至关重要。本研究开发了一种检测范围广、灵敏度高的利妥昔单抗（rituximab-TRFIA）的时间分辨荧光免疫分析法（TRFIA），旨在加强MN治疗中的治疗药物监测。方法：采用链霉亲和素包被微孔板、生物素化抗利妥昔单抗独特型抗体和Eu3+标记的靶向利妥昔单抗Fc片段的小鼠抗人IgG，制备捕获型利妥昔单抗- trfia。该试验用于测量接受利妥昔单抗治疗的MN患者的利妥昔单抗血清浓度。结果：利妥昔单抗- trfia在0.50 ~ 2500 ng/mL线性范围内，检出限（LOD）为0.062 ng/mL。内变异系数（CV）为1.97 % ~ 9.50 %，间变异系数为7.44 % ~ 9.99 %。回收率为99.14 % ~ 107.75 %。与其他单克隆抗体（mab）无交叉反应。检测范围比酶联免疫吸附试验（ELISA）提高2个数量级，灵敏度提高6倍。与ELISA相比，Rituximab-TRFIA在同一测量范围内具有较强的一致性（P < 0.0001）。MN患者血清利妥昔单抗水平显著高于对照组（P < 0.0001）。在未达到缓解的患者中，60% %至70% %的患者药物浓度不足，利妥昔单抗药代动力学符合预期趋势。结论：rituximab- trfia为监测MN患者血清rituximab浓度提供了高灵敏度、宽检测范围和可靠的性能，支持个性化治疗策略。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文去求助

来源期刊

Clinica Chimica Acta 医学-医学实验技术

CiteScore

10.10

自引率

2.00%

发文量

1268

审稿时长

23 days

期刊介绍： The Official Journal of the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) Clinica Chimica Acta is a high-quality journal which publishes original Research Communications in the field of clinical chemistry and laboratory medicine, defined as the diagnostic application of chemistry, biochemistry, immunochemistry, biochemical aspects of hematology, toxicology, and molecular biology to the study of human disease in body fluids and cells. The objective of the journal is to publish novel information leading to a better understanding of biological mechanisms of human diseases, their prevention, diagnosis, and patient management. Reports of an applied clinical character are also welcome. Papers concerned with normal metabolic processes or with constituents of normal cells or body fluids, such as reports of experimental or clinical studies in animals, are only considered when they are clearly and directly relevant to human disease. Evaluation of commercial products have a low priority for publication, unless they are novel or represent a technological breakthrough. Studies dealing with effects of drugs and natural products and studies dealing with the redox status in various diseases are not within the journal''s scope. Development and evaluation of novel analytical methodologies where applicable to diagnostic clinical chemistry and laboratory medicine, including point-of-care testing, and topics on laboratory management and informatics will also be considered. Studies focused on emerging diagnostic technologies and (big) data analysis procedures including digitalization, mobile Health, and artificial Intelligence applied to Laboratory Medicine are also of interest.