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基于液体油泥的蛋白质电动浓缩。

Liquid Plasticine-Based Electrokinetic Enrichment of Proteins.

机构信息

The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, 710049, Xi'an, P. R. China.

Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, 710049, Xi'an, P. R. China.

出版信息

ChemistryOpen. 2023 Mar;12(3):e202200259. doi: 10.1002/open.202200259.

Abstract

Protein analysis is an important approach for disease diagnosis, in which sample pretreatment is an essential step since protein samples are often complex and many protein biomarkers are of low abundance. Here, given the good openness and light transmission of liquid plasticine (LP), which is a liquid entity formed by SiO nanoparticles and encapsulated aqueous solution, we developed a LP-based field-amplified sample stacking (FASS) system for protein enrichment. The system was composed of a LP container, a sample solution and a Tris-HCl solution containing hydroxyethyl cellulose (HEC). The system design, mechanism investigation, optimization of experimental parameters and characterization of LP-FASS performance for protein enrichment were well studied. Under the optimized experimental conditions of 1 % HEC, 100 mm Tris-HCl and 100 V in the LP-FASS system, a 40-80 times enrichment of proteins was obtained in 40 min using bovine hemoglobin (BHb) as the model protein using the constructed LP-FASS system. The simultaneous enrichment of multiple proteins (phycocyanin, BHb and cytochrome C) was also realized using the system. The LP-FASS system can serve as a new platform for protein enrichment which is easy to be combined with online and offline detections.

摘要

蛋白质分析是疾病诊断的重要方法,其中样品预处理是必不可少的步骤,因为蛋白质样品通常很复杂,并且许多蛋白质生物标志物的丰度较低。在这里,考虑到液体塑料(LP)具有良好的开放性和透光性,它是由 SiO 纳米粒子和封装水溶液形成的液体实体,我们开发了一种基于 LP 的场放大样品堆积(FASS)系统用于蛋白质富集。该系统由 LP 容器、样品溶液和含有羟乙基纤维素(HEC)的 Tris-HCl 溶液组成。对该系统的设计、机制研究、实验参数优化以及 LP-FASS 对蛋白质富集性能的表征进行了深入研究。在 LP-FASS 系统中 1%HEC、100mm Tris-HCl 和 100V 的优化实验条件下,使用构建的 LP-FASS 系统,以牛血红蛋白(BHb)为模型蛋白,在 40min 内可获得 40-80 倍的蛋白质富集。该系统还可实现多种蛋白质(藻蓝蛋白、BHb 和细胞色素 C)的同时富集。LP-FASS 系统可作为一种新的蛋白质富集平台,易于与在线和离线检测相结合。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd6b/10041546/2c051e329b4e/OPEN-12-e202200259-g005.jpg

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