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Into the theory of the partial‐filling affinity capillary electrophoresis and the determination of apparent stability constants of analyte‐ligand complexes
Author(s) -
Ansorge Martin,
Dubský Pavel,
Ušelová Kateřina
Publication year - 2018
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.201700385
Subject(s) - analyte , capillary electrophoresis , ligand (biochemistry) , chemistry , affinity electrophoresis , electrophoresis , linearization , stability (learning theory) , analytical chemistry (journal) , electrolyte , chromatography , affinity chromatography , computer science , nonlinear system , electrode , physics , receptor , enzyme , biochemistry , quantum mechanics , machine learning
The partial‐filling affinity capillary electrophoresis (pf‐ACE) works with a ligand present in a background electrolyte that forms a weak complex with an analyte. In contrast to a more popular mobility‐shift affinity capillary electrophoresis, only a short plug of the ligand is introduced into a capillary in the pf‐ACE. Both methods can serve for determining apparent stability constants of the formed complexes but this task is hindered in the pf‐ACE by the fact that the analyte spends only a part of its migration time in a contact with the ligand. In 1998, Amini and Westerlund published a linearization strategy that allows for extracting an effective mobility of an analyte in the presence of a neutral ligand out of the pf‐ACE data. The main purpose of this paper is to show that the original formula is only approximate. We derive a new formula and demonstrate its applicability by means of computer simulations. We further inspect several strategies of data processing in the pf‐ACE regarding a risk of an error propagation. This establishes a good practice of determining apparent stability constants of analyte‐ligand complexes by means of the pf‐ACE.