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Separation and zeta-potential determination of proteins and their oligomers using electrical asymmetrical flow field-flow fractionation (EAF4)

Choi, Jaeyeong LU orcid ; Fuentes, Catalina LU ; Fransson, Jonas ; Wahlgren, Marie LU orcid and Nilsson, Lars LU (2020) In Journal of Chromatography A 1633.
Abstract

Electrical asymmetrical flow field-flow fractionation (EAF4) is an interesting new analytical technique that separates proteins based on size or molecular weight and simultaneously determines the electrical characteristics of each population. However, until now, the research using EAF4 has not been published except for the proof-of-concept in the original publication by Johann et. al. in 2015 [1]. Hence the methods capabilities and optimized conditions need to be further investigated, such as composition of the carrier liquid, pH stability and effect of the electric field strength. The pH instability was observed in the initial method of EAF4 due to the electrolysis products when applied electric field. Therefore, we have investigated... (More)

Electrical asymmetrical flow field-flow fractionation (EAF4) is an interesting new analytical technique that separates proteins based on size or molecular weight and simultaneously determines the electrical characteristics of each population. However, until now, the research using EAF4 has not been published except for the proof-of-concept in the original publication by Johann et. al. in 2015 [1]. Hence the methods capabilities and optimized conditions need to be further investigated, such as composition of the carrier liquid, pH stability and effect of the electric field strength. The pH instability was observed in the initial method of EAF4 due to the electrolysis products when applied electric field. Therefore, we have investigated and provided a modified method for rapid pH stabilization through additional focusing step with the electric field. Then, the electrical properties such as the zeta-potential and effective net charge of the monomer and oligomers of three different proteins (GA-Z, BSA, and Ferritin) were determined based on their electrophoretic mobility from EAF4. The results showed that there were limitations to the applicability of separation by EAF4 to proteins. Nevertheless, this study shows that EAF4 is an interesting new technique that can examine the zeta-potential of individual proteins in mixtures (or monomers and oligomers) not accessible by other techniques.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Effective net charge, Electrical asymmetrical flow field-flow fractionation (EAF4), Electrical characteristics, Proteins, Separation, Zeta-potential
in
Journal of Chromatography A
volume
1633
article number
461625
publisher
Elsevier
external identifiers
  • scopus:85093658261
  • pmid:33128976
ISSN
0021-9673
DOI
10.1016/j.chroma.2020.461625
language
English
LU publication?
yes
id
01bf1159-379e-4145-a238-96d3f900cc1d
date added to LUP
2020-11-04 11:35:43
date last changed
2024-04-03 14:07:57
@article{01bf1159-379e-4145-a238-96d3f900cc1d,
  abstract     = {{<p>Electrical asymmetrical flow field-flow fractionation (EAF4) is an interesting new analytical technique that separates proteins based on size or molecular weight and simultaneously determines the electrical characteristics of each population. However, until now, the research using EAF4 has not been published except for the proof-of-concept in the original publication by Johann et. al. in 2015 [1]. Hence the methods capabilities and optimized conditions need to be further investigated, such as composition of the carrier liquid, pH stability and effect of the electric field strength. The pH instability was observed in the initial method of EAF4 due to the electrolysis products when applied electric field. Therefore, we have investigated and provided a modified method for rapid pH stabilization through additional focusing step with the electric field. Then, the electrical properties such as the zeta-potential and effective net charge of the monomer and oligomers of three different proteins (GA-Z, BSA, and Ferritin) were determined based on their electrophoretic mobility from EAF4. The results showed that there were limitations to the applicability of separation by EAF4 to proteins. Nevertheless, this study shows that EAF4 is an interesting new technique that can examine the zeta-potential of individual proteins in mixtures (or monomers and oligomers) not accessible by other techniques.</p>}},
  author       = {{Choi, Jaeyeong and Fuentes, Catalina and Fransson, Jonas and Wahlgren, Marie and Nilsson, Lars}},
  issn         = {{0021-9673}},
  keywords     = {{Effective net charge; Electrical asymmetrical flow field-flow fractionation (EAF4); Electrical characteristics; Proteins; Separation; Zeta-potential}},
  language     = {{eng}},
  publisher    = {{Elsevier}},
  series       = {{Journal of Chromatography A}},
  title        = {{Separation and zeta-potential determination of proteins and their oligomers using electrical asymmetrical flow field-flow fractionation (EAF4)}},
  url          = {{http://dx.doi.org/10.1016/j.chroma.2020.461625}},
  doi          = {{10.1016/j.chroma.2020.461625}},
  volume       = {{1633}},
  year         = {{2020}},
}