Design Guidelines of Free-Flow Counterflow Gradient Focusing Device for Protein Fractionation.

IF 3 3区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

ELECTROPHORESIS Pub Date : 2025-02-18 DOI:10.1002/elps.8106

Matthew Courtney, Oscar Manuel Carreno-Molina, Carolyn L Ren

引用次数: 0

Abstract

Free-flow counterflow gradient focusing (FF-CGF) is a promising tool for protein fractionation. This work establishes guidelines to construct an FF-CGF system suitable for preparative applications. Numerical and analytical computations are undertaken to investigate design parameters, including device geometry, flow conditions, sample concentration, and applied voltage. This collective information serves as a design tool to guide the fabrication of the device, which, in this case, is constructed from polymethyl methacrylate (PMMA) and double-sided tape. Furthermore, the design tool also supports device operation and provides insights into the expected resolution and throughput. Experimental results, which demonstrated the ability of the designed chip to generate a uniform counterflow gradient while separating small molecules and proteins, validated the accuracy of the design guidelines.

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蛋白质分离用自由流动逆流梯度聚焦装置的设计指南。

自由流动逆流梯度聚焦（FF-CGF）是一种很有前途的蛋白质分离工具。本工作为构建适合制备应用的FF-CGF体系建立了指导方针。采用数值和解析计算来研究设计参数，包括器件几何形状、流动条件、样品浓度和施加电压。这些集体信息可以作为指导设备制造的设计工具，在这种情况下，该设备由聚甲基丙烯酸甲酯（PMMA）和双面胶带构成。此外，设计工具还支持设备操作，并提供对预期分辨率和吞吐量的见解。实验结果表明，设计的芯片能够在分离小分子和蛋白质时产生均匀的逆流梯度，验证了设计指南的准确性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

ELECTROPHORESIS 生物-分析化学

CiteScore

6.30

自引率

13.80%

发文量

244

审稿时长

1.9 months

期刊介绍： ELECTROPHORESIS is an international journal that publishes original manuscripts on all aspects of electrophoresis, and liquid phase separations (e.g., HPLC, micro- and nano-LC, UHPLC, micro- and nano-fluidics, liquid-phase micro-extractions, etc.). Topics include new or improved analytical and preparative methods, sample preparation, development of theory, and innovative applications of electrophoretic and liquid phase separations methods in the study of nucleic acids, proteins, carbohydrates natural products, pharmaceuticals, food analysis, environmental species and other compounds of importance to the life sciences. Papers in the areas of microfluidics and proteomics, which are not limited to electrophoresis-based methods, will also be accepted for publication. Contributions focused on hyphenated and omics techniques are also of interest. Proteomics is within the scope, if related to its fundamentals and new technical approaches. Proteomics applications are only considered in particular cases. Papers describing the application of standard electrophoretic methods will not be considered. Papers on nanoanalysis intended for publication in ELECTROPHORESIS should focus on one or more of the following topics: • Nanoscale electrokinetics and phenomena related to electric double layer and/or confinement in nano-sized geometry • Single cell and subcellular analysis • Nanosensors and ultrasensitive detection aspects (e.g., involving quantum dots, "nanoelectrodes" or nanospray MS) • Nanoscale/nanopore DNA sequencing (next generation sequencing) • Micro- and nanoscale sample preparation • Nanoparticles and cells analyses by dielectrophoresis • Separation-based analysis using nanoparticles, nanotubes and nanowires.