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首页> 外文期刊>Animal Science Journal >Analysis of differential strategies to enhance detection of low-abundance proteins in the bovine serum proteome
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Analysis of differential strategies to enhance detection of low-abundance proteins in the bovine serum proteome

机译：鉴别策略提高牛血清蛋白质中低丰度蛋白质的差异策略分析

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摘要

Serum-based biomarkers hold propitious applications for addressing livestock health, and management. However, discovery of protein biomarkers in complex biological fluids like serum is wholly intractable due to the large dynamic range of protein concentrations; that is, similar to 10-12 high abundance proteins constitute >90% of the total protein content and effectively mask proteomic detection of low-abundance biomarkers. Toward addressing this limitation, we test a continuous elution size-based fractionation method, and two approaches that use affinity interaction-based separation of proteins in preparing bovine serum, and compare liquid chromatography tandem mass spectrometry protein identification to neat serum. Our results identify the high-abundance proteins in bovine serum, and demonstrate dynamic range compression and improved protein identification with the different enrichment methods. Although these findings indicate the highest protein number identified in bovine serum (445 proteins, all methods combined), and by any single sample processing method (312 proteins) to date, they still remain lower than levels deemed necessary for biomarker discovery. As such, this investigation revealed limitations to resolving the bovine serum proteome, and the need for species-specific tools for immunodepleting high-abundance proteins. In concert, this study represents a step toward advancing sample preparation methods for bovine serum biomarker identification.
机译：基于血清的生物标志物可持有卓越的应用，以解决牲畜健康和管理。然而，在复杂的生物流体中发现蛋白质生物标志物，如血清的血清，由于蛋白质浓度的巨大动态范围而完全难以解决;也就是说，类似于10-12个高丰度蛋白质，占总蛋白质含量的90％，有效掩盖低丰度生物标志物的蛋白质组学检测。为了解决这一限制，我们测试一种基于连续的洗脱尺寸的分馏方法，以及在制备牛血清中使用基于亲和相互作用的蛋白质分离的两种方法，并将液相色谱串联质谱蛋白鉴定与整齐的血清相比。我们的结果鉴定了牛血清中的高丰度蛋白，并表明了具有不同富集方法的动态范围压缩和改善的蛋白质鉴定。虽然这些发现表明牛血清中鉴定的最高蛋白质数（445个蛋白，所有方法组合），并且通过任何单一样品加工方法（312蛋白）迄今为止，它们仍然低于生物标志物发现所需的水平。因此，该研究揭示了解决牛血清蛋白质组的局限性，以及用于免疫凝固蛋白的物种特异性工具。在音乐会中，该研究代表了推进牛血清生物标志物鉴定的样品制备方法的步骤。

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