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Mid‐scale free‐flow electrophoresis with gravity‐induced uniform flow of background buffer in chamber for the separation of cells and proteins
Author(s) -
Dong YuChao,
Shao Jing,
Yin XiaoYang,
Fan Liuyin,
Cao ChengXi
Publication year - 2011
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201100293
Subject(s) - chromatography , chemistry , electrophoresis , analytical chemistry (journal) , tris , buffer (optical fiber) , bovine serum albumin , biochemistry , telecommunications , computer science
A large‐scale free‐flow electrophoresis (LS‐FFE) is often too large for cell separation of lab scale, whereas micro‐FFE (μFFE) has great difficulty in cell isolation due to easy blockage by cell accumulation in μFFE. In this study, a mid‐scale FFE (MS‐FFE) is developed for cell and protein separations. The volume of the separation chamber (70×40×0.1–0.8 mm) is from 280 μL to 2.24 mL, much lower than that in an LS‐FFE but higher than that in a μFFE. Gravity is used for uniform flow of the background buffer only via a single pump with 16 channels and the sample is injected via an adjuster originally used for clinical intravenous injection. The experiments reveal that the hydrodynamic and electrohydrodynamic flows are much stable, and the Joule heat can be effectively dispersed without obvious positive or negative deviation as shown by the omega plots. By the device, Escherichia coli and Staphylococcus aureus , which easily accumulate to block μFFE and are separated with difficulty due to their same negative charges carried, can be well isolated under the conditions of 4.5 mM pH 8.5 Tris‐boric buffer (4.5 mM Tris, 4.5 mM boric acid) with 0.10 mM ethylene diamine tetraacetic acid and 5% m/v sucrose, 200 μL/min, 800 V, and sample injection via inlet 4. The mid‐scale FFE device could also be used for the separation of three model proteins of horse heart cytochrome c, myoglobin and bovine serum albumin. The device has clear significance for mid‐scale separation of cells and proteins.