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Separation of mAbs Molecular Variants by Analytical HIC-HPLC

Published: Thursday, June 05, 2014
Last Updated: Thursday, June 05, 2014
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HIC-HPLC is a less widely used analytical technique compared with standard methods for separating variants based on size, charge or electrophoretic mobility.

Hydrophobic interaction chromatography-high performance liquid chromatography (HIC-HPLC) is a powerful analytical method used for the separation of molecular variants of therapeutic proteins. The method has been employed for monitoring various post-translational modifications, including proteolytic fragments and domain misfolding in etanercept (Enbrel®); tryptophan oxidation, aspartic acid isomerization, the formation of cyclic imide, and α amidated carboxy terminus in recombinant therapeutic monoclonal antibodies; and carboxy terminal heterogeneity and serine fucosylation in Fc and Fab fragments. HIC-HPLC is also a powerful analytical technique for the analysis of antibody-drug conjugates. Most current analytical columns, methods, and applications are described, and critical method parameters and suitability for operation in regulated environment are discussed, in this review

The review article, Separation of mAbs molecular variants by analytical hydrophobic interaction chromatography HPLC: overview and applications, is published online in Landes Bioscience and is free to access. 

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