Protein purification by off-gel electrophoresis

Alexandra Ros, Michel Faupel, Hervé Mees, Jan Van Oostrum, Rosaria Ferrigno, Frédéric Reymond, Philippe Michel, Joël S. Rossier, Hubert H. Girault

Research output: Contribution to journalArticle

94 Citations (Scopus)

Abstract

A novel free-flow protein purification technique based on isoelectric electrophoresis is presented, where the proteins are purified in solution without the need of carrier ampholytes. The gist of the method is to flow protein solutions under an immobilised pH gradient gel (IPG) through which an electric field is applied perpendicular to the direction of the flow. Due to the buffering capacity of the IPG gel, proteins with an isoelectric point (p/) close to pH of the gel in contact with the flow chamber stay in solution because they are neutral and therefore not extracted by the electric field. Other proteins will be charged when approaching the IPG gel and are extracted into the gel by the electric field. Both a demonstration experiment with p/ markers and a simulation of the electric field distribution are presented to highlight the principle of the system. In addition, an isoelectric fractionation of an Escherichia coil extract is shown to illustrate the possible applications.

Original languageEnglish (US)
Pages (from-to)151-156
Number of pages6
JournalProteomics
Volume2
Issue number2
DOIs
StatePublished - 2002
Externally publishedYes

Fingerprint

Electrophoresis
Purification
Gels
Proton-Motive Force
Electric fields
Proteins
Ampholyte Mixtures
Escherichia
Isoelectric Point
Fractionation
Contacts (fluid mechanics)
Demonstrations
Experiments

Keywords

  • Isoelectric focusing
  • Isoelectric purification
  • Prefractionation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Ros, A., Faupel, M., Mees, H., Van Oostrum, J., Ferrigno, R., Reymond, F., ... Girault, H. H. (2002). Protein purification by off-gel electrophoresis. Proteomics, 2(2), 151-156. https://doi.org/10.1002/1615-9861(200202)2:2<151::AID-PROT151>3.0.CO;2-9

Protein purification by off-gel electrophoresis. / Ros, Alexandra; Faupel, Michel; Mees, Hervé; Van Oostrum, Jan; Ferrigno, Rosaria; Reymond, Frédéric; Michel, Philippe; Rossier, Joël S.; Girault, Hubert H.

In: Proteomics, Vol. 2, No. 2, 2002, p. 151-156.

Research output: Contribution to journalArticle

Ros, A, Faupel, M, Mees, H, Van Oostrum, J, Ferrigno, R, Reymond, F, Michel, P, Rossier, JS & Girault, HH 2002, 'Protein purification by off-gel electrophoresis', Proteomics, vol. 2, no. 2, pp. 151-156. https://doi.org/10.1002/1615-9861(200202)2:2<151::AID-PROT151>3.0.CO;2-9
Ros, Alexandra ; Faupel, Michel ; Mees, Hervé ; Van Oostrum, Jan ; Ferrigno, Rosaria ; Reymond, Frédéric ; Michel, Philippe ; Rossier, Joël S. ; Girault, Hubert H. / Protein purification by off-gel electrophoresis. In: Proteomics. 2002 ; Vol. 2, No. 2. pp. 151-156.
@article{039da41859b14054a52e07e5747b0df3,
title = "Protein purification by off-gel electrophoresis",
abstract = "A novel free-flow protein purification technique based on isoelectric electrophoresis is presented, where the proteins are purified in solution without the need of carrier ampholytes. The gist of the method is to flow protein solutions under an immobilised pH gradient gel (IPG) through which an electric field is applied perpendicular to the direction of the flow. Due to the buffering capacity of the IPG gel, proteins with an isoelectric point (p/) close to pH of the gel in contact with the flow chamber stay in solution because they are neutral and therefore not extracted by the electric field. Other proteins will be charged when approaching the IPG gel and are extracted into the gel by the electric field. Both a demonstration experiment with p/ markers and a simulation of the electric field distribution are presented to highlight the principle of the system. In addition, an isoelectric fractionation of an Escherichia coil extract is shown to illustrate the possible applications.",
keywords = "Isoelectric focusing, Isoelectric purification, Prefractionation",
author = "Alexandra Ros and Michel Faupel and Herv{\'e} Mees and {Van Oostrum}, Jan and Rosaria Ferrigno and Fr{\'e}d{\'e}ric Reymond and Philippe Michel and Rossier, {Jo{\"e}l S.} and Girault, {Hubert H.}",
year = "2002",
doi = "10.1002/1615-9861(200202)2:2<151::AID-PROT151>3.0.CO;2-9",
language = "English (US)",
volume = "2",
pages = "151--156",
journal = "Proteomics",
issn = "1615-9853",
publisher = "Wiley-VCH Verlag",
number = "2",

}

TY - JOUR

T1 - Protein purification by off-gel electrophoresis

AU - Ros, Alexandra

AU - Faupel, Michel

AU - Mees, Hervé

AU - Van Oostrum, Jan

AU - Ferrigno, Rosaria

AU - Reymond, Frédéric

AU - Michel, Philippe

AU - Rossier, Joël S.

AU - Girault, Hubert H.

PY - 2002

Y1 - 2002

N2 - A novel free-flow protein purification technique based on isoelectric electrophoresis is presented, where the proteins are purified in solution without the need of carrier ampholytes. The gist of the method is to flow protein solutions under an immobilised pH gradient gel (IPG) through which an electric field is applied perpendicular to the direction of the flow. Due to the buffering capacity of the IPG gel, proteins with an isoelectric point (p/) close to pH of the gel in contact with the flow chamber stay in solution because they are neutral and therefore not extracted by the electric field. Other proteins will be charged when approaching the IPG gel and are extracted into the gel by the electric field. Both a demonstration experiment with p/ markers and a simulation of the electric field distribution are presented to highlight the principle of the system. In addition, an isoelectric fractionation of an Escherichia coil extract is shown to illustrate the possible applications.

AB - A novel free-flow protein purification technique based on isoelectric electrophoresis is presented, where the proteins are purified in solution without the need of carrier ampholytes. The gist of the method is to flow protein solutions under an immobilised pH gradient gel (IPG) through which an electric field is applied perpendicular to the direction of the flow. Due to the buffering capacity of the IPG gel, proteins with an isoelectric point (p/) close to pH of the gel in contact with the flow chamber stay in solution because they are neutral and therefore not extracted by the electric field. Other proteins will be charged when approaching the IPG gel and are extracted into the gel by the electric field. Both a demonstration experiment with p/ markers and a simulation of the electric field distribution are presented to highlight the principle of the system. In addition, an isoelectric fractionation of an Escherichia coil extract is shown to illustrate the possible applications.

KW - Isoelectric focusing

KW - Isoelectric purification

KW - Prefractionation

UR - http://www.scopus.com/inward/record.url?scp=0036211004&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036211004&partnerID=8YFLogxK

U2 - 10.1002/1615-9861(200202)2:2<151::AID-PROT151>3.0.CO;2-9

DO - 10.1002/1615-9861(200202)2:2<151::AID-PROT151>3.0.CO;2-9

M3 - Article

VL - 2

SP - 151

EP - 156

JO - Proteomics

JF - Proteomics

SN - 1615-9853

IS - 2

ER -