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Protein separation by electrophoresis in a nonsieving amphoteric medium
Author(s) -
Blanco Stéphane,
Clifton Michael J.,
Joly JeanLouis,
Peltre Gabriel
Publication year - 1996
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150170624
Subject(s) - isoelectric point , isoelectric focusing , conductivity , chemistry , electrophoresis , separation process , resolution (logic) , buffer (optical fiber) , chromatography , protein purification , separation (statistics) , analytical chemistry (journal) , high resolution , biochemistry , computer science , telecommunications , remote sensing , artificial intelligence , machine learning , enzyme , geology
A numerical model has been used to study the separation of protein mixtures by zone electrophoresis in a nonsieving amphoteric medium. An amphoteric buffer fixes the pH of the solution close to its isoelectric point, where the buffer molecules are uncharged: they thus contribute very little to the conductivity of the solution. This means that high field strengths can be used for rapid separation without sacrificing resolution. The numerical study shows that in this process the band spreading that can reduce resolution is essentially due to differences in migration rate between protein‐rich and low‐protein zones: both differences in solution conductivity and in protein mobility are involved. Rules for judging the buffer capacity of amphoteric molecules are presented and it is shown how, for a given protein, the effectiveness of this technique varies with the range of pH in which it is applied.