Bringing an immuno-PCR-based pharmacodynamic biomarker assay for reduced human IL-33 in serum, plasma, and aqueous humor into the clinic

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-08-07 DOI:10.1016/j.jpba.2025.117095

John Hok Nin Lowe , Trung Nguy , Randall Dere , Vahan B. Indjeian , Mauricio Maia

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引用次数: 0

Abstract

Immuno-polymerase chain reaction (iPCR) is an analytical technology that combines the specificity of antibody reagents with the sensitivity of polymerase chain reaction (PCR) signal amplification. Here we describe the optimization and qualification of an ultra-sensitive iPCR method designed to detect and quantify active (reduced form) interleukin-33 (IL-33) in human samples. The modified assay incorporates several improvements over a previous version utilized in research studies. The changes aimed at making the assay more suitable to the rigorous requirements promulgated to biomarker assays supporting clinical development of new human therapeutics. The assay can accurately measure sub-picogram/mL levels of reduced IL-33, and has a dynamic range of 977–1000,000 fg/mL. We found this iPCR-based format to be versatile, and we believe it can be adapted to detect and quantify other low-level cytokines in human matrices when limited sample volumes are available.

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将基于免疫pcr的人类血清、血浆和房水中IL-33减少的药效学生物标志物测定方法引入临床

免疫聚合酶链反应（immune -polymerase chain reaction, iPCR）是将抗体试剂的特异性与聚合酶链反应（polymerase chain reaction， PCR）信号扩增的敏感性相结合的分析技术。在这里，我们描述了一种超灵敏iPCR方法的优化和鉴定，该方法设计用于检测和定量人类样本中活性（还原形式）白介素-33 （IL-33）。修改后的测定法结合了在研究中使用的先前版本的几个改进。这些变化旨在使测定更适合于支持新人类疗法临床开发的生物标志物测定的严格要求。该方法可准确测定IL-33还原亚图/mL水平，动态范围为977 - 1000000 fg/mL。我们发现这种基于ipcr的格式是通用的，我们相信它可以在有限的样本量下用于检测和量化人类基质中的其他低水平细胞因子。

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.