title: Separation of mAbs molecular variants by analytical hydrophobic interaction chromatography HPLC: overview and applications.
abstract: 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.
authors.forename: Mark
authors.initials: M
authors.lastname: Haverick
authors.forename: Selina
authors.initials: S
authors.lastname: Mengisen
authors.forename: Mohammed
authors.initials: M
authors.lastname: Shameem
authors.forename: Alexandre
authors.initials: A
authors.lastname: Ambrogelly
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006031:Glycosylation; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007127:Immunoglobulin Constant Regions / Q000737:chemistry / Q000302:isolation & purification*; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D007127:Immunoglobulin Constant Regions; D007140:Immunoglobulin Fab Fragments; D011994:Recombinant Proteins
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