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Rapid analysis of charge variants of monoclonal antibodies with capillary zone electrophoresis in dynamically coated fused‐silica capillary
Author(s) -
He Yan,
Isele Colleen,
Hou Weiying,
Ruesch Margaret
Publication year - 2011
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201000719
Subject(s) - capillary electrophoresis , capillary action , chromatography , capillary electrochromatography , monoclonal antibody , chemistry , electrophoresis , analytical chemistry (journal) , materials science , antibody , biology , composite material , immunology
A capillary zone electrophoresis (CZE) method was developed for the rapid analysis of charge heterogeneity of immunoglobulin G (IgG) monoclonal antibodies (mAbs). The separation was carried out in a short, dynamically coated fused‐silica capillary. A number of separation parameters were investigated and optimized, including pH, concentration of the separation buffer (ε‐amino caproic acid), concentration of the triethylenetetramine (TETA) dynamic coating, the capillary internal diameter and the field strength used for the separation. The effects of between‐run flushing of the capillary and the data acquisition rate were also evaluated. Under the optimized conditions, a fast (<5 min), selective and reproducible separation of mAb charge variants was achieved under a very high electric field strength (1000 V/cm). This method also requires only a short conditioning of the capillary, with between‐run conditioning completed within 2 min. The method was evaluated for specificity, sensitivity, linearity, accuracy and precision. The same separation conditions were applied to the rapid separation (2–5 min) of charge variants of multiple monoclonal antibodies with p I in the range of 7.0–9.5. Compared with other existing methods for charge variants analysis, this method has several advantages including a short run time, rapid capillary conditioning and simple sample preparation.