Profiling post-centrifugation delay of serum and plasma with antibody bead arrays

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• 作者：Ulrika Qundos ; Mun-Gwan Hong ; Gunnel Tybring ; Mark Divers ; Jacob Odeberg ; Mathias Uhlen ; Peter Nilsson ; Jochen M. Schwenk
• 关键词：FDR ; false discovery rate ; MFI ; median fluorescence intensity ; HPA ; Human Protein Atlas ; nMFI ; normalized median fluorescence intensity ; LMM ; linear mixed effects model ; CV ; coefficient of variance.
• 刊名：Journal of Proteomics
• 出版年：16 December, 2013
• 年：2013
• 卷：95
• 期：Complete
• 页码：46-54
• 全文大小：677 K

文摘

Several biobanking initiatives have emerged to create extensive collections of specimen for biomedical studies and various analytical platforms. An affinity proteomic analysis with antibody suspension bead arrays was conducted to investigate the influence of the pre-analytical time and temperature conditions on blood derived samples. Serum and EDTA plasma prepared from 16 individuals was centrifuged and aliquots were kept either at 4 掳C or in ambient temperature for 1 h and up to 36 h prior to first storage. Multiplexed protein profiles of post-centrifugation delay were generated in 384 biotinylated samples using 373 antibodies that targeted 343 unique proteins. Very few profiles were observed as significantly altered by the studied temperature and time intervals. Single binder and sandwich assays revealed decreasing levels of caldesmon 1 (CALD1) related to EDTA standard tubes and prolonged post-centrifugation delay of 36 h. Indications from changes in CALD1 levels require further confirmation in independent material, but the current data suggests that samples should preferentially be frozen during the day of collection when to be profiled with antibody arrays selected for this study.

Biological significance

Affinity-based profiling of serum and plasma by microarray assays can provide unique opportunities for the discovery of biomarkers. It is though often not known how differences in sample handling after collection influence the downstream analysis. By profiling three types of blood preparations for alterations in protein profiles with respect to time and temperature post centrifugation, we addressed an important component in the analysis and of such specimen. We believe that this analysis adds valuable information to be considered when biobanking blood derived samples.

This article is part of a Special Issue entitled: Standardization and Quality Control in Proteomics.