Peripheral blood mononuclear cells isolation from mouse and rabbit blood to quantify active nucleotide and define brincidofovir dose for smallpox.

IF 1.9 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Bioanalysis Pub Date : 2024-10-30 DOI:10.1080/17576180.2024.2418245

Robert Hart, Shane Karnik, Kathy Van Sickle, Mark Mullin, Odin Naderer, Tim Tippin, John Dunn

引用次数: 0

Abstract

Aim: Brincidofovir has been approved by the US FDA for the treatment of smallpox via the "Animal Rule". The active moiety, cidofovir diphosphate (CDV-PP), was measurable in human peripheral blood mononuclear cells (PBMCs), but quantitation in animals was challenging given their limited blood volume. The aim of this study was to optimize PBMC isolation from rabbit and mouse blood to allow quantitation of CDV-PP.Materials & methods: PBMC isolation methods using various separation media were evaluated using blood from naive and brincidofovir-dosed animals. CDV-PP was quantified using liquid chromatography tandem mass spectrometry.Results & conclusion: PBMC isolation yields were increased by >fourfold compared with yields obtained using human methods, allowing cross-species exposure comparisons.

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从小鼠和家兔血液中分离外周血单核细胞，以量化活性核苷酸并确定治疗天花的布林昔多福韦剂量。

目的：美国食品和药物管理局通过 "动物规则 "批准布林昔多福韦用于治疗天花。活性分子西多福韦二磷酸酯（CDV-PP）可在人类外周血单核细胞（PBMC）中测量，但由于动物的血容量有限，在动物体内进行定量分析具有挑战性。本研究的目的是优化兔和小鼠血液中 PBMC 的分离，以便对 CDV-PP 进行定量：材料与方法：使用天真动物和布林昔多福韦剂量动物的血液，对使用各种分离介质的 PBMC 分离方法进行了评估。采用液相色谱串联质谱法对 CDV-PP 进行定量：结果与结论：与人类方法相比，PBMC 分离率提高了四倍以上，因此可以进行跨物种暴露比较。

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

Bioanalysis BIOCHEMICAL RESEARCH METHODS-CHEMISTRY, ANALYTICAL

CiteScore

3.30

自引率

16.70%

发文量

审稿时长

2 months

期刊介绍： Reliable data obtained from selective, sensitive and reproducible analysis of xenobiotics and biotics in biological samples is a fundamental and crucial part of every successful drug development program. The same principles can also apply to many other areas of research such as forensic science, toxicology and sports doping testing. The bioanalytical field incorporates sophisticated techniques linking sample preparation and advanced separations with MS and NMR detection systems, automation and robotics. Standards set by regulatory bodies regarding method development and validation increasingly define the boundaries between speed and quality. Bioanalysis is a progressive discipline for which the future holds many exciting opportunities to further reduce sample volumes, analysis cost and environmental impact, as well as to improve sensitivity, specificity, accuracy, efficiency, assay throughput, data quality, data handling and processing. The journal Bioanalysis focuses on the techniques and methods used for the detection or quantitative study of analytes in human or animal biological samples. Bioanalysis encourages the submission of articles describing forward-looking applications, including biosensors, microfluidics, miniaturized analytical devices, and new hyphenated and multi-dimensional techniques. Bioanalysis delivers essential information in concise, at-a-glance article formats. Key advances in the field are reported and analyzed by international experts, providing an authoritative but accessible forum for the modern bioanalyst.