de Morais-Zani Karen, Grego Kathleen Fernandes, Tanaka Aparecida Sadae, Tanaka-Azevedo Anita Mitico
Laborato´rios de Fisiopatologia, Instituto Butantan, São Paulo, Brazil.
J Biomol Tech. 2011 Jul;22(2):67-73.
The proteomic analysis of plasma samples represents a challenge as a result of the presence of highly abundant proteins such as albumin. To enable the detection of biomarkers, which are commonly low-abundance proteins, in complex blood fluids, it is necessary to remove high-abundance proteins efficiently. Moreover, there is a range of about 10 orders of magnitude for the abundance of different protein species in serum. Here, we describe for the first time a study of reptilian albumin depletion using resins usually used in mammalian plasma depletion procedures. We performed the depletion of albumin from Bothrops jaraca plasma using the HiTrap Blue high-performance column (GE Healthcare Life Sciences, Piscataway, NJ, USA) and the kit Albumin & IgG Depletion SpinTrap column (GE Healthcare Life Sciences). In addition, proteomic approaches were used to analyze reptilian plasma. Our results showed that B. jararaca albumin bound to both columns, but those interactions were not enough to remove a large amount of albumin to reach an enrichment of low-abundance proteins. Although the depletion techniques used in this work were not the best to remove B. jararaca plasma albumin, our present work highlights the similarity between B. jararaca and mammalian albumin, contributing to the knowledge of comparative hemostatic proteins.
由于存在诸如白蛋白等高度丰富的蛋白质,血浆样本的蛋白质组学分析面临挑战。为了能够在复杂的血液流体中检测通常为低丰度蛋白质的生物标志物,有效去除高丰度蛋白质是必要的。此外,血清中不同蛋白质种类的丰度范围约为10个数量级。在此,我们首次描述了一项使用通常用于哺乳动物血浆去除程序的树脂进行爬行动物白蛋白去除的研究。我们使用HiTrap Blue高性能柱(美国新泽西州皮斯卡塔韦的通用电气医疗生命科学公司)和白蛋白及IgG去除SpinTrap柱试剂盒(通用电气医疗生命科学公司)从具窍蝮蛇血浆中去除白蛋白。此外,采用蛋白质组学方法分析爬行动物血浆。我们的结果表明,具窍蝮蛇白蛋白与两种柱都结合,但这些相互作用不足以去除大量白蛋白以实现低丰度蛋白质的富集。尽管本研究中使用的去除技术并非去除具窍蝮蛇血浆白蛋白的最佳方法,但我们目前的工作突出了具窍蝮蛇与哺乳动物白蛋白之间的相似性,有助于对比较止血蛋白的了解。