应用随机寡核苷酸文库修饰磁性颗粒提高血浆蛋白质的鉴定覆盖度

发布时间：2018-01-27 04:52

本文关键词： 随机寡核苷酸文库血浆蛋白质高丰度低丰度谱图数量　出处：《色谱》2017年03期 　论文类型：期刊论文

【摘要】：基于随机寡核苷酸与蛋白质之间的离子、亲和、疏水、氢键等相互作用力及多种空间结构作用,发展了一种基于随机寡核苷酸文库作为配基的新型血浆样品处理方法。采用寡核苷酸文库修饰的磁性颗粒(MNP@ssDNA)材料,在生理缓冲体系条件下捕获血浆样品中的蛋白质。比较了两种洗脱体系的洗脱效果,并利用nano-RPLC-ESIMS/MS对获得的蛋白质酶解液组分进行分析。结果表明,MNP@ssDNA材料处理后的血浆蛋白质鉴定数量提升了约29.5%,两种洗脱体系呈现良好的互补性(26.7%)。血浆中前10种高丰度蛋白质的谱图占有率从处理前的31.82%降低到21.31%(洗脱体系1)和26.20%(洗脱体系2)。在鉴定到的蛋白质中,丰度最低的蛋白质在血浆中的质量浓度约为0.29 ng/mL,该蛋白质仅在MNP@ssDNA材料处理后被鉴定到。结果证明MNP@ssDNA策略不仅能有效降低血浆中高丰度蛋白质的丰度,也为低丰度蛋白质的深度挖掘提供了新的思路。
[Abstract]:Based on the interaction between random oligonucleotide and protein, such as ion, affinity, hydrophobic, hydrogen bond and so on, as well as a variety of spatial structures. A novel plasma sample processing method based on random oligonucleotide library was developed. The protein in plasma samples was captured under physiological buffer system. The elution effects of two elution systems were compared. Nano-RPLC-ESIMS/MS was used to analyze the components of protein hydrolysate. The number of identified plasma proteins increased by about 29.5% after treatment with MNP@ssDNA materials. The two elution systems showed good complementarities. The percentage of the first 10 kinds of high abundance protein in plasma decreased from 31.82% before treatment to 21.31% (elution system 1). And 26.20% (elution system 2) in identified proteins. The lowest concentration of protein in plasma was about 0.29 ng/mL. The protein was identified only after MNP@ssDNA treatment. The results showed that MNP@ssDNA strategy could not only effectively reduce the abundance of high abundance protein in plasma. It also provides a new idea for deep mining of low abundance protein.
【作者单位】：中国烟草总公司郑州烟草研究院;中国科学院大连化学物理研究所中科院分离分析化学重点实验室国家色谱研究分析中心;郑州大学第一附属医院医学研究中心;
【基金】：国家自然科学基金项目(21505158,81600046)~~
【分类号】：R392-33
【正文快照】： *通讯联系人.Tel:(0411)84379720,E-mail:lihuazhang@dicp.ac.cn.血浆在生命科学及临床医学领域具有重要的研究价值,但血浆内蛋白质的组成复杂,浓度动态范围宽。据文献[1,2]报道,血浆中前10种高丰度蛋白质占蛋白质总含量的90%,前22种高丰度蛋白质占蛋白质总含量的99%,而血浆中

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