Comparative Evaluation of Smart Sampling for hCG Determination: A New Potential Direction in Protein Biomarker Analysis From Dried Microsamples.

IF 2.7 3区 医学 Q2 BIOCHEMICAL RESEARCH METHODS
Ago Mrsa, Marijana Matijevic, Yvette Dehnes, Trine Grønhaug Halvorsen, Léon Reubsaet
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Abstract

Since the early 20th century, sampling biological matrices like blood on paper (dried blood spots [DBS]) has been vital in clinical analysis. While DBS microsampling is convenient for small molecules, extensive sample preparation can make LC-MS protein analysis impractical because of the time-consuming steps, especially for low-abundance proteins. Smart sampling, introduced in 2018, simplifies this by integrating sample preparation directly on the sampler. The work presented in this paper aims to compare a newly validated smart sampling method with two other methods: an in-house method based on immunocapture on magnetic beads and a commercial method that uses electrochemiluminescence immunoassay (ECLIA). The performance of the three hCG methods was compared using 21 single-blind serum samples. Linear regression analysis revealed strong correlations (all R2 > 0.91) between the actual sample concentrations and the results obtained from all three methods. Immunocapture with magnetic beads showed the strongest linear correlation (R2 = 0.974). To assess agreement between the methods, Bland-Altman analysis was conducted. The comparison between smart sampling and magnetic beads showed an average bias of -5.2, with no significant trend in variation across the sample concentration range of 0.5-75 ng/mL. The smart sampling and ECLIA comparison revealed a bias of 0.4 ± 4 ng/mL, indicating even better agreement and consistent results. This paper presents the first-ever comparison of a smart sampling method with existing methods. The results highlight smart sampling as a promising new approach for bioanalysis and boost the technique as a viable alternative in protein biomarker analysis from complex matrices using LC-MS.

智能取样测定hCG的比较评价:干燥微样品中蛋白质生物标志物分析的新方向。
自20世纪初以来,对纸上血液(干血斑[DBS])等生物基质取样在临床分析中起着至关重要的作用。虽然DBS微采样对小分子很方便,但大量的样品制备可能使LC-MS蛋白质分析变得不切实际,因为步骤耗时,特别是对于低丰度的蛋白质。2018年推出的智能采样通过直接在采样器上集成样品制备来简化这一过程。本文提出的工作旨在将新验证的智能采样方法与其他两种方法进行比较:基于磁珠免疫捕获的内部方法和使用电化学发光免疫测定(ECLIA)的商业方法。用21份单盲血清样本比较3种hCG方法的性能。线性回归分析显示,实际样品浓度与三种方法所得结果之间存在很强的相关性(R2均为> 0.91)。磁珠免疫捕获法的线性相关性最强(R2 = 0.974)。为了评估方法之间的一致性,进行了Bland-Altman分析。智能采样与磁珠的平均偏差为-5.2,在0.5-75 ng/mL的样品浓度范围内没有明显的变化趋势。智能取样和ECLIA比较显示偏差为0.4±4 ng/mL,表明结果更加一致和一致。本文首次将智能采样方法与现有方法进行了比较。结果强调智能采样是一种有前途的生物分析新方法,并将该技术作为使用LC-MS分析复杂基质中蛋白质生物标志物的可行替代方法。
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来源期刊
Drug Testing and Analysis
Drug Testing and Analysis BIOCHEMICAL RESEARCH METHODS-CHEMISTRY, ANALYTICAL
CiteScore
5.90
自引率
24.10%
发文量
191
审稿时长
2.3 months
期刊介绍: As the incidence of drugs escalates in 21st century living, their detection and analysis have become increasingly important. Sport, the workplace, crime investigation, homeland security, the pharmaceutical industry and the environment are just some of the high profile arenas in which analytical testing has provided an important investigative tool for uncovering the presence of extraneous substances. In addition to the usual publishing fare of primary research articles, case reports and letters, Drug Testing and Analysis offers a unique combination of; ‘How to’ material such as ‘Tutorials’ and ‘Reviews’, Speculative pieces (‘Commentaries’ and ‘Perspectives'', providing a broader scientific and social context to the aspects of analytical testing), ‘Annual banned substance reviews’ (delivering a critical evaluation of the methods used in the characterization of established and newly outlawed compounds). Rather than focus on the application of a single technique, Drug Testing and Analysis employs a unique multidisciplinary approach to the field of controversial compound determination. Papers discussing chromatography, mass spectrometry, immunological approaches, 1D/2D gel electrophoresis, to name just a few select methods, are welcomed where their application is related to any of the six key topics listed below.
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