两种尿蛋白组成分析方法在肾脏疾病诊断及随访中的比较。

IF 2.5 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS
ELECTROPHORESIS Pub Date : 2025-09-20 DOI:10.1002/elps.70037
Joris Guyon, Jeanne Malaubier, Hong-Van Pham, Lila Rami Arab, Laurence Pieroni, Marie-Laure Curutchet-Burtin, Rémi Segues, Sylvie Caspar-Bauguil, Annie M Bérard
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引用次数: 0

摘要

蛋白尿分析对于在肾小球滤过率恶化之前发现肾脏疾病的早期阶段和监测治疗后肾脏疾病的进展是必要的。电泳通常是第一个定向测试,尽管这个测试是耗时的,其解释可能是主观的。两种类型的尿蛋白电泳凝胶可用:(1)高分辨率(HR)琼脂糖凝胶和(2)琼脂糖凝胶结合免疫检测特异性尿蛋白电泳迁移(UP)。由于前者是单克隆蛋白定量的最佳方法,后者是单克隆蛋白表征的最佳方法,因此我们在研究中只研究了确定肾损伤类型的方法。因此,我们的研究目的是比较两种蛋白尿分型的策略:UP凝胶和HR凝胶,可以量化特定的蛋白质(白蛋白,最终是转铁蛋白,α1-微球蛋白,最终是β2-微球蛋白,免疫球蛋白G (IgG), α2-巨球蛋白,如果有血液存在),HR凝胶可以可视化RBP, β2-微球蛋白和转铁蛋白。两种方法在检测肾小管标志物(α1-微球蛋白和β2-微球蛋白)、非选择性肾小球标志物(IgG)和肾后标志物(α2-巨球蛋白)排泄异常方面具有可比性。白蛋白的检测限比免疫法定量限低25倍,结果不同,导致假阳性,无法区分尿白蛋白的低排泄和高排泄。综上所述,对于UP凝胶,当在不检查单克隆蛋白的情况下进行蛋白尿分型时,我们建议对特定蛋白(如白蛋白、α1微球蛋白、最终是β2微球蛋白和IgG)进行免疫测定。在相关单克隆蛋白研究的背景下,小管和肾小球损伤标志物(不包括白蛋白)可以在UP凝胶上解释。如果血液存在,α2-巨球蛋白必须用免疫分析法测定α2-巨球蛋白/白蛋白的比值,以确定蛋白尿的肾后起源。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Comparison of Two Strategies of Analysis of Urinary Protein Composition for the Diagnosis and Follow-Up of Renal Diseases.

Proteinuria analysis is necessary to detect the early stages of kidney disease before the estimated glomerular filtration rate deteriorates and to monitor the progression of treated kidney disease. Electrophoresis is often the first orientation test, although this test is time-consuming and its interpretation may be subjective. Two types of electrophoresis gel for urinary proteins are available: (1) high-resolution (HR) agarose gel and (2) agarose gel combined with immunological detection of specific urinary proteins after their electrophoretic migration (UP). As the former is known to provide the best results for the quantification of monoclonal protein and the latter for its characterization, we only investigated methods for determining the type of kidney damage in our study. Therefore, the aim of our study was to compare two strategies for proteinuria typing: UP gel to HR gel with quantification of specific proteins (albumin, eventually transferrin, α1-microglobulin, eventually β2-microglobulin, immunoglobulin G (IgG), and α2-macroglobulin if blood is present), and HR gel allowing the visualization of RBP, β2-microglobulin, and transferrin. The two methods were comparable in detecting abnormal excretion of tubular markers (α1-microglobulin and β2-microglobulin), nonselective glomerular markers such as IgG, and post-renal marker (α2-macroglobulin). The results differed for albumin, whose limit of detection was 25 times lower than the limit of quantification by immunoassay, leading to false positives and no differentiation between low and high excretion of albuminuria. In conclusion on UP gel, when proteinuria typing is prescribed without investigating monoclonal protein, we recommend carrying out immunoassays for specific proteins (e.g., albumin, α1-microglobulin, eventually β2-microglobulin, and IgG). In a context of associated monoclonal protein investigation, tubular and glomerular damage markers (excluding albumin) can be interpreted on a UP gel. If blood is present, α2-macroglobulin must be measured using immunoassays to determine the post-renal origin of proteinuria using the ratio α2-macroglobulin/albumin.

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来源期刊
ELECTROPHORESIS
ELECTROPHORESIS 生物-分析化学
CiteScore
6.30
自引率
13.80%
发文量
244
审稿时长
1.9 months
期刊介绍: ELECTROPHORESIS is an international journal that publishes original manuscripts on all aspects of electrophoresis, and liquid phase separations (e.g., HPLC, micro- and nano-LC, UHPLC, micro- and nano-fluidics, liquid-phase micro-extractions, etc.). Topics include new or improved analytical and preparative methods, sample preparation, development of theory, and innovative applications of electrophoretic and liquid phase separations methods in the study of nucleic acids, proteins, carbohydrates natural products, pharmaceuticals, food analysis, environmental species and other compounds of importance to the life sciences. Papers in the areas of microfluidics and proteomics, which are not limited to electrophoresis-based methods, will also be accepted for publication. Contributions focused on hyphenated and omics techniques are also of interest. Proteomics is within the scope, if related to its fundamentals and new technical approaches. Proteomics applications are only considered in particular cases. Papers describing the application of standard electrophoretic methods will not be considered. Papers on nanoanalysis intended for publication in ELECTROPHORESIS should focus on one or more of the following topics: • Nanoscale electrokinetics and phenomena related to electric double layer and/or confinement in nano-sized geometry • Single cell and subcellular analysis • Nanosensors and ultrasensitive detection aspects (e.g., involving quantum dots, "nanoelectrodes" or nanospray MS) • Nanoscale/nanopore DNA sequencing (next generation sequencing) • Micro- and nanoscale sample preparation • Nanoparticles and cells analyses by dielectrophoresis • Separation-based analysis using nanoparticles, nanotubes and nanowires.
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