Campbell Amanda J, Palstrøm Nicolai B, Rasmussen Lars M, Lindholt Jes S, Beck Hans C
Department of Clinical Biochemistry, Odense University Hospital, Odense, Denmark.
Center for Clinical Proteomics (CCP), Odense University Hospital, Odense, Denmark.
Clin Proteomics. 2025 May 18;22(1):20. doi: 10.1186/s12014-025-09540-w.
Microsamples are simple blood sampling procedures utilizing small blood draws. Although microsamples are regularly used in some disciplines, proteomic analysis of these samples is an emerging field. Currently, it is unclear whether the quantitative precision and proteome coverage achieved in microsamples is comparable to plasma or serum. As a consequence, microsamples are not used in proteomics to the same degree as more traditional blood samples.
The objective of this scoping review was to report the applications of microsamples within clinical mass spectrometry-based proteomics. This was accomplished by describing both proof-of-concept and clinical proteomics research within this field, with an additional evaluation of the newest advances regarding clinical proteomics.
Original scientific literature was included where bottom-up mass spectrometry was used to analyze endogenous proteins from human microsamples.
Relevant publications were sourced through three scientific databases (MEDLINE, EMBASE and Scopus) in addition to backward and forward citation searches through Scopus. Record screening was performed independently by two separate authors. The review was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Extension for Scoping Reviews (PRISMA-ScR) guidelines.
A total of 209 records were screened for inclusion from database searches and 3157 records were screened from forward and backward citation searches, resulting in 64 eligible studies. An evaluation of proof-of-concept research within this field revealed that although microsamples are amenable to high-throughput proteomics using a variety of targeted and untargeted acquisition methods, quantification remained a relevant issue. Microsampling practices were heterogeneous, and no standard procedure existed for protein quantification. Clinical studies investigated protein expression in numerous disease or experimental groups, including hemoglobinopathies and immunodeficiency disorders.
The use of microsamples is increasing within the proteomics field and these samples are amenable to standard bottom-up workflows. Although microsamples present a clear advantage in terms of sampling procedure, both the sample collection and quantification procedures remain to be standardized. However, there is an incentive to address the remaining issues, since microsampling would greatly reduce the resources necessary to sample large cohorts within clinical proteomics, a field that currently lacks large discovery and validation cohorts.
微量样本是利用少量采血进行的简单血液采样程序。尽管微量样本在某些学科中经常使用，但对这些样本进行蛋白质组分析仍是一个新兴领域。目前，尚不清楚微量样本中实现的定量精度和蛋白质组覆盖范围是否与血浆或血清相当。因此，微量样本在蛋白质组学中的使用程度不及更传统的血液样本。
本范围综述的目的是报告微量样本在基于临床质谱的蛋白质组学中的应用。这是通过描述该领域的概念验证和临床蛋白质组学研究，并对临床蛋白质组学的最新进展进行额外评估来实现的。
纳入使用自下而上质谱法分析人类微量样本中内源性蛋白质的原始科学文献。
除了通过Scopus进行的前后引文检索外，还通过三个科学数据库（MEDLINE、EMBASE和Scopus）获取相关出版物。记录筛选由两位独立作者独立进行。本综述按照系统评价和Meta分析扩展的范围综述的首选报告项目（PRISMA-ScR）指南进行。
通过数据库检索共筛选出209条记录以纳入研究，通过前后引文检索筛选出3157条记录，最终有64项符合条件的研究。对该领域概念验证研究的评估表明，尽管微量样本适用于使用各种靶向和非靶向采集方法的高通量蛋白质组学，但定量仍然是一个相关问题。微量采样方法各不相同，蛋白质定量没有标准程序。临床研究调查了众多疾病或实验组中的蛋白质表达，包括血红蛋白病和免疫缺陷疾病。
微量样本在蛋白质组学领域的使用正在增加，并且这些样本适用于标准的自下而上工作流程。尽管微量样本在采样程序方面具有明显优势，但样本采集和定量程序仍有待标准化。然而，解决剩余问题是有动力的，因为微量采样将大大减少临床蛋白质组学中对大型队列进行采样所需的资源，而该领域目前缺乏大型的发现和验证队列。
