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Analysis of recombinant monoclonal antibodies by capillary zone electrophoresis
Author(s) -
Espinosa de la Garza Carlos E.,
PerdomoAbúndez Francisco C.,
PadillaCalderón Jesús,
UribeWiechers Jaime M.,
Pérez Néstor O.,
FloresOrtiz Luis F.,
MedinaRivero Emilio
Publication year - 2013
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.201200575
Subject(s) - electropherogram , repeatability , capillary electrophoresis , chromatography , chemistry , monoclonal antibody , gene isoform , recombinant dna , comparability , computational biology , analytical chemistry (journal) , antibody , biology , biochemistry , immunology , mathematics , combinatorics , gene
Analytical platforms that characterize charge heterogeneity in therapeutic proteins, such as m A bs, are important tools that can be used to define quality attributes. CZE separates protein moieties close to their native state and is a valuable physicochemical analytical method that can be used in parallel with other orthogonal methods for characterization and comparability. In this study, custom conditions for the analysis of charge heterogeneity of two m A bs were developed with regard to critical parameters in the BGE , running conditions, and sample treatment. The method application was tested for up to four m A bs and one m A b fragment. The electropherograms showed specific profiles and contrasting levels of basic and acidic isoforms with respect to the main isoform. Issues that surround this method, such as peak tailing and capillary lifetime, are summarized. Using this method, the identities of rituximab and trastuzumab were confirmed, based on the correspondence between the biosimilars and reference products, noninterference of the sample matrix, and the ability to separate spiked samples of related m A bs. The RSD of the isoform content and migration time for the method repeatability were less than 2 and 1%, respectively.