Simultaneous separation of negatively and positively charged species in dynamic field gradient focusing using a dual polarity electric field

Jeffrey M Burke; Zheng Huang; Cornelius F Ivory

doi:10.1021/ac901634y

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Simultaneous separation of negatively and positively charged species in dynamic field gradient focusing using a dual polarity electric field

Journal article

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Peer reviewed

Simultaneous separation of negatively and positively charged species in dynamic field gradient focusing using a dual polarity electric field

Jeffrey M Burke, Zheng Huang and Cornelius F Ivory

Analytical chemistry (Washington), Vol.81(19), pp.8236-8243

10/01/2009

DOI: https://doi.org/10.1021/ac901634y

Handle:

https://hdl.handle.net/2376/108647

PMCID: PMC3236110

PMID: 19722517

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https://doi.org/10.1021/ac901634yView

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Abstract

Cytochromes c - isolation & purification

Electrodes

Isoelectric Focusing - instrumentation

Isoelectric Focusing - methods

Phycoerythrin - isolation & purification

Hydrogen-Ion Concentration

Dynamic field gradient focusing (DFGF) utilizes an electric field gradient established by a computer-controlled electrode array to separate and concentrate charged analytes at unique axial positions. Traditionally, DFGF has been restricted to the analysis of negatively charged species due to limitations in the software of our voltage controller. This paper introduces a new voltage controller capable of operating under normal polarity (positive potentials applied to the electrode array) and reversed polarity (negative potentials applied to the electrode array) for the separation of negatively and positively charged analytes, respectively. The experiments conducted under normal polarity and reversed polarity illustrate the utility of the new controller to perform reproducible DFGF separations (elution times showing less than 1% run-to-run variation) over a wide pH range (3.08 to 8.5) regardless of the protein charge. A dual polarity experiment is then shown in which the separation channel has been divided into normal polarity and reversed polarity regions. This simultaneous separation of negatively charged R-phycoerythrin (R-PE) and positively charged cytochrome c (CYTC) within the same DFGF apparatus is shown.

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Title: Simultaneous separation of negatively and positively charged species in dynamic field gradient focusing using a dual polarity electric field
Creators: Jeffrey M Burke - Gene and Linda Voiland School of Chemical Engineering and Bioengineering, Washington State University, Pullman, WA, USA
Zheng Huang
Cornelius F Ivory
Publication Details: Analytical chemistry (Washington), Vol.81(19), pp.8236-8243
Academic Unit: Chemical Engineering and Bioengineering, School of
Publisher: United States
Grant note: T32 GM008336-18 / NIGMS NIH HHS T32 GM008336 / NIGMS NIH HHS TM2GM08336 / NIGMS NIH HHS T32 GM008336-20 / NIGMS NIH HHS T32 GM008336-19 / NIGMS NIH HHS
Identifiers: 99900547639801842
Language: English
Resource Type: Journal article