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Loading capacity of carrier ampholytes – based buffers in capillary electrophoresis
Author(s) -
Busnel JeanMarc,
Descroix Stéphanie,
Godfrin Dominique,
Hennion MarieClaire,
Peltre Gabriel
Publication year - 2006
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.200500646
Subject(s) - isoelectric focusing , chemistry , chromatography , electrophoresis , ionic strength , isoelectric point , analytical chemistry (journal) , electric field , aqueous solution , biochemistry , physics , quantum mechanics , enzyme
Narrow pH cuts of carrier ampholytes (CAs), originally designed for IEF, have been used as BGEs in CE. Their physicochemical properties, rather high buffering capacity and low conductivity, allow very efficient protein separations under high electric field strength. Due to their isoelectric properties, CA BGEs are expected to present a low ionic concentration and consequently a low loading capacity. In this study, we developed a simple method that allows the estimation of the loading capacity of a UV‐absorbing BGE by CE. We first characterized in terms of loading capacity, classical ammediol‐chromate UV‐absorbing BGEs and a 10 mM histidine solution, a classical isoelectric buffer. Then, the loading capacity of four different CA‐based BGEs has been assessed. Experimental results have shown that the CA‐based buffers were presenting a rather high loading capacity, comparable to classical buffer ones and far higher than the one of the 10 mM histidine solution.