title: Quantitation of soluble aggregates in recombinant monoclonal antibody cell culture by pH-gradient protein A chromatography.
abstract: Monoclonal antibodies (mAbs) produced from mammalian cell culture may contain significant amounts of dimers and higher order aggregates. Quantitation of soluble aggregates in the cell culture is time-consuming and labor-intensive, usually involving a purification step to remove the impurities that interfere with the subsequent size exclusion chromatography (SEC) analysis. We have developed a novel pH-gradient protein A chromatography for rapid, non-size based separation of the aggregates in mAb cell culture samples. Our results demonstrate that this method has excellent correlation with SEC and can be applied to both human immunoglobulin gamma 1 (IgG1) and IgG2 antibodies. This approach can be useful in the quantitation of soluble aggregates in crude cell culture samples.
authors.affiliation: Analytical and Formulation Sciences, One Amgen Center Drive, Thousand Oaks, CA 91320, USA. hpan@amgen.com
authors.forename: Hai
authors.initials: H
authors.lastname: Pan
authors.forename: Ken
authors.initials: K
authors.lastname: Chen
authors.forename: Matt
authors.initials: M
authors.lastname: Pulisic
authors.forename: Izydor
authors.initials: I
authors.lastname: Apostol
authors.forename: Gang
authors.initials: G
authors.lastname: Huang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D016466:CHO Cells; D002846:Chromatography, Affinity / Q000379:methods*; D002850:Chromatography, Gel; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D013205:Staphylococcal Protein A / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D013205:Staphylococcal Protein A