Impact of hemolysis on the levels of proteins associated with aging and age-related neurodegenerative diseases in a multicentric clinical research

IF 1.3 Q3 MEDICAL LABORATORY TECHNOLOGY Practical Laboratory Medicine Pub Date : 2025-04-01 Epub Date: 2025-01-20 DOI:10.1016/j.plabm.2025.e00455

Masroor Anwar , Km Renu , Abhinay Kumar Singh , Abhilasha Nayal , Bharat Thyagarajan , Peifeng Hu , Jinkook Lee , Sharmistha Dey , A.B. Dey

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Abstract

Introduction

Hemolysis is a known interference factor that has been found to show erroneous effect. Present study analyzes the impact of hemolysis on the concentrations of protein biomarkers of Alzheimer's disease (Aβ42, t-Tau, p-Tau181) along with novel proteins which are currently under investigation (SIRT1,SIRT2,SIRT6,FOXO3A, NFL, Aβ40, GFAP).

Methods

Plasma samples were grouped into two categories: hemolyzed and non-hemolyzed groups. Degree of hemolysis (in percentage) was separately analyzed using Single molecule array (SIMOA) technology. Quantitative analysis for hemolyzed and non-hemolyzed samples were done using surface plasmon resonance (SPR) technology.

Results

The SIMOA analysis indicated that at high levels of hemolysis (1000 mg/dL) there was an increase in NFL protein level up to approximately 30 % whereas p-Tau181 did not show much interference even at higher hemolysate concentration. Aβ40, Aβ42 and GFAP showed modest effect up to hemolysis of 250mg/dL-500 mg/dL. SPR analysis of total Tau (t-Tau), p-Tau181, SIRT1, SIRT6 showed the consistency in the result and there was no significant difference in hemolyzed plasma compared to non-hemolyzed samples. Aβ42 and FOXO3A showed decline in hemolyzed plasma compared to non-hemolyzed samples (4.34 ± 0.18ng/ul; 4.95 ± 0.19ng/ul) and (3.83 ± 0.34ng/ul; 5.12 ± 0.46ng/ul), respectively whereas, a significant increase in the concentration was observed for SIRT2; 2.4 ± 0.10ng/ul in hemolyzed compared to 1.30 ± 0.22ng/ul in non-hemolyzed group.

Conclusions

High grade hemolysis leads to altered protein concentration associated with neurodegeneration. Present study emphasizes the need to have pre-analytical inspection for hemolysis detection especially in a multicentric biomarker study.

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一项多中心临床研究中溶血对与衰老和年龄相关的神经退行性疾病相关蛋白水平的影响

溶血是一种已知的干扰因素，已被发现显示出错误的效果。本研究分析了溶血对阿尔茨海默病蛋白生物标志物（a - β42、t-Tau、p-Tau181）以及目前正在研究的新蛋白（SIRT1、SIRT2、SIRT6、FOXO3A、NFL、a - β40、GFAP）浓度的影响。方法将血浆分为溶血组和非溶血组。采用单分子阵列（SIMOA）技术分别分析溶血程度（百分比）。采用表面等离子体共振（SPR）技术对溶血和非溶血样品进行定量分析。结果SIMOA分析表明，在高溶血水平（1000 mg/dL）下，NFL蛋白水平增加约30%，而p-Tau181即使在高溶血浓度下也没有表现出太大的干扰。a - β40、a - β42和GFAP在溶血剂量为250mg/dL- 500mg /dL时，溶血效果一般。总Tau蛋白（t-Tau）、p-Tau181、SIRT1、SIRT6的SPR分析结果一致，溶血血浆与非溶血血浆无显著差异。溶血血浆中Aβ42和FOXO3A较未溶血血浆下降(4.34±0.18ng/ul；4.95±0.19ng/ul)和（3.83±0.34ng/ul）；5.12±0.46ng/ul)，而SIRT2的浓度显著增加；溶血组为2.4±0.10ng/ul，非溶血组为1.30±0.22ng/ul。结论高度溶血导致蛋白浓度改变与神经退行性变有关。目前的研究强调需要有溶血检测的分析前检查，特别是在一个多中心的生物标志物研究。

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

Practical Laboratory Medicine Health Professions-Radiological and Ultrasound Technology

CiteScore

3.50

自引率

0.00%

发文量

审稿时长

7 weeks

期刊介绍： Practical Laboratory Medicine is a high-quality, peer-reviewed, international open-access journal publishing original research, new methods and critical evaluations, case reports and short papers in the fields of clinical chemistry and laboratory medicine. The objective of the journal is to provide practical information of immediate relevance to workers in clinical laboratories. The primary scope of the journal covers clinical chemistry, hematology, molecular biology and genetics relevant to laboratory medicine, microbiology, immunology, therapeutic drug monitoring and toxicology, laboratory management and informatics. We welcome papers which describe critical evaluations of biomarkers and their role in the diagnosis and treatment of clinically significant disease, validation of commercial and in-house IVD methods, method comparisons, interference reports, the development of new reagents and reference materials, reference range studies and regulatory compliance reports. Manuscripts describing the development of new methods applicable to laboratory medicine (including point-of-care testing) are particularly encouraged, even if preliminary or small scale.