自行加工的非对称流场流分离仪器对蛋白质分离的应用研究

发布时间：2018-01-16 22:24

本文关键词：自行加工的非对称流场流分离仪器对蛋白质分离的应用研究　出处：《中国药学杂志》2016年16期 　论文类型：期刊论文

【摘要】：目的设计并加工一套针对生物大分子分离的非对称流场流分离(asymmetric flow field-flow fractionation,As Fl FFF或AF4)仪器,对仪器性能进行初步研究,为下一步实际样品的分离提供依据和参考经验。方法为了解决样品聚焦(focus)问题,自行研究了一种新的聚焦方式。以标准牛血清白蛋白和乙醇脱氢酶作为混合样品,对仪器性能进行验证。并通过改变进样/聚焦时间、横向流动速度(cross-in flow)和横向流出速度(cross-out flow),考察各因素对分离效果(分离度R表示)的影响。结果混合蛋白的分离度分别随聚焦时间的减少和横向流动速度的增加而增加,而横向流出速度的影响并不显著。结论该非对称流场流分离仪器已基本实现对混合蛋白的分离,预计可用于更为复杂的样品分离。
[Abstract]:Objective to design and process a set of asymmetric flow field-flow fractionation for the separation of biological macromolecules. In order to solve the problem of focusing on samples, the performance of as F1 FFF or AF4 instrument is studied preliminarily, which provides the basis and reference experience for the separation of practical samples in the next step. A new focusing method was studied. The standard bovine serum albumin (BSA) and ethanol dehydrogenase (EDH) were used as mixed samples to verify the performance of the instrument, and the injection / focus time was changed. The lateral flow velocity was cross-flow) and the lateral outflow velocity was cross-out. The results showed that the separation degree of mixed protein increased with the decrease of focusing time and the increase of lateral flow velocity. Conclusion the asymmetric flow separation instrument has basically achieved the separation of mixed proteins, which can be used for more complex sample separation.
【作者单位】：北京大学药学院;中国农业大学;
【分类号】：TH79
【正文快照】： 生物大分子的分离表征一直是困扰研究者的重要问题,现在常用的分子生物学手段均无法实现温和、快速、高效的分离,存在样品前处理复杂,分离时间长,尺寸分辨率低,对高分子量颗粒无法分离等缺点[1-2]。本实验开发的非对称流场流分离仪器能分离和表征几纳米至数十微米的颗粒,可弥

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