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用于生物样本库目的的唾液样本蛋白质质量评估

Protein Quality Assessment on Saliva Samples for Biobanking Purposes.

作者信息

Rosa Nuno, Marques Jéssica, Esteves Eduardo, Fernandes Mónica, Mendes Vera M, Afonso Ângela, Dias Sérgio, Pereira Joaquim Polido, Manadas Bruno, Correia Maria José, Barros Marlene

机构信息

1 Institute of Health Sciences (ICS), Center for Interdisciplinary Research in Health (CIIS), Universidade Católica Portuguesa , Viseu, Portugal .

2 Department of Biology, University of Aveiro , Aveiro, Portugal .

出版信息

Biopreserv Biobank. 2016 Aug;14(4):289-97. doi: 10.1089/bio.2015.0054. Epub 2016 Mar 3.

Abstract

Biobank saliva sample quality depends on specific criteria applied to collection, processing, and storage. In spite of the growing interest in saliva as a diagnostic fluid, few biobanks currently store large collections of such samples. The development of a standard operating procedure (SOP) for saliva collection and quality control is fundamental for the establishment of a new saliva biobank, which stores samples to be made available to the saliva research community. Different collection methods were tested regarding total volume of protein obtained, protein content, and protein profiles, and the results were used to choose the best method for protein studies. Furthermore, the impact of the circadian variability and inter- and intraindividual differences, as well as the saliva sample stability at room temperature, were also evaluated. Considering our results, a sublingual cotton roll method for saliva collection proved to produce saliva with the best characteristics and should be applied in the morning, whenever possible. In addition, there is more variability in salivary proteins between individuals than in the same individual for a 5-month period. According to the electrophoretic protein profile, protein stability is guaranteed for 24 hours at room temperature and the protein degradation profile and protein identification were characterized. All this information was used to establish an SOP for saliva collection, processing, and storage in a biobank. We conclude that it is possible to collect saliva using an easy and inexpensive protocol, resulting in saliva samples for protein analysis with sufficient quality for biobanking purposes.

摘要

生物样本库唾液样本的质量取决于采集、处理和储存所适用的特定标准。尽管人们对唾液作为一种诊断液的兴趣日益浓厚,但目前很少有生物样本库储存大量此类样本。制定唾液采集和质量控制的标准操作规程(SOP)对于建立一个新的唾液生物样本库至关重要,该样本库储存的样本可供唾液研究界使用。针对获得的蛋白质总量、蛋白质含量和蛋白质谱测试了不同的采集方法,并将结果用于选择蛋白质研究的最佳方法。此外,还评估了昼夜节律变异性、个体间和个体内差异的影响,以及唾液样本在室温下的稳定性。根据我们的结果,舌下棉卷法采集的唾液具有最佳特性,应尽可能在早晨应用。此外,个体间唾液蛋白质的变异性比同一个体在5个月内的变异性更大。根据电泳蛋白质谱,蛋白质在室温下24小时内稳定性良好,并对蛋白质降解谱和蛋白质鉴定进行了表征。所有这些信息都被用于建立生物样本库中唾液采集、处理和储存的SOP。我们得出结论,使用简单且廉价的方案采集唾液是可行的,从而获得用于蛋白质分析的唾液样本,其质量足以满足生物样本库的目的。

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