Department of Proteomics, School of Biotechnology, KTH - Royal Institute of Technology, Albanova University Center, Stockholm, Sweden.
Proteomics. 2010 Feb;10(3):532-40. doi: 10.1002/pmic.200900657.
In the pursuit towards a systematic analysis of human diseases, array-based approaches within antibody proteomics offer high-throughput strategies to discover protein biomarkers in serum and plasma. To investigate the influence of sample preparation on such discovery attempts, we report on a systematic effort to compare serum and plasma protein profiles determined with an antibody suspension bead array. The intensity levels were used to define protein profiles and no significant differences between serum and plasma were observed for 79% of the 174 antibodies (targeting 156 proteins). By excluding 36 antibodies giving rise to differential intensity levels, cluster analysis revealed donor-specific rather than preparation-dependent grouping. With a cohort from a clinically relevant medical condition, the metabolic syndrome, the influence of the sample type on a multiplexed biomarker discovery approach was further investigated. Independent comparisons of protein profiles in serum and plasma revealed an antibody targeting ADAMTSL-4, a protein that would qualify to be studied further in association with the condition. In general, the preparation type had an impact on the results of the applied antibody suspension bead array, and while the technical variability was equal, plasma offered a greater biological variability and allowed to give rise to more discoveries than serum.
在对人类疾病进行系统分析的过程中，抗体蛋白质组学中的基于阵列的方法为在血清和血浆中发现蛋白质生物标志物提供了高通量的策略。为了研究样品制备对这种发现尝试的影响，我们报告了一项系统的研究，以比较使用抗体悬浮珠阵列确定的血清和血浆蛋白质谱。强度水平用于定义蛋白质谱，并且在 174 种抗体（针对 156 种蛋白质）中，79%的血清和血浆之间没有观察到显著差异。通过排除导致差异强度水平的 36 种抗体，聚类分析显示出供体特异性而非制备依赖性分组。对于来自临床相关医学病症（代谢综合征）的队列，进一步研究了样品类型对多路复用生物标志物发现方法的影响。血清和血浆中蛋白质谱的独立比较揭示了一种针对 ADAMTSL-4 的抗体，该蛋白有望进一步研究与该病症的关联。一般来说，制备类型对应用的抗体悬浮珠阵列的结果有影响，虽然技术变异性相等，但血浆提供了更大的生物学变异性，并允许产生比血清更多的发现。
