title: Modeling on-column reduction of trisulfide bonds in monoclonal antibodies during protein A chromatography.
abstract: Trisulfides can be a common post-translational modification in many recombinant monoclonal antibodies. These are a source of product heterogeneity that add to the complexity of product characterization and hence, need to be reduced for consistent product quality. Trisulfide bonds can be converted to the regular disulfide bonds by incorporating a novel cysteine wash step during Protein A affinity chromatography. An empirical model is developed for this on-column reduction reaction to compare the reaction rates as a function of typical operating parameters such as temperature, cysteine concentration, reaction time and starting level of trisulfides. The model presented here is anticipated to assist in the development of optimal wash conditions for the Protein A step to effectively reduce trisulfides to desired levels.
authors.affiliation: Process Biochemistry, Technical Development, Biogen, Research Triangle Park, NC, United States. Electronic address: sanchayita.ghose@biogen.com.
authors.forename: Sanchayita
authors.initials: S
authors.lastname: Ghose
authors.affiliation: Process Biochemistry, Technical Development, Biogen, Research Triangle Park, NC, United States.
authors.forename: Rupshika
authors.initials: R
authors.lastname: Rajshekaran
authors.affiliation: Process Biochemistry, Technical Development, Biogen, Research Triangle Park, NC, United States.
authors.forename: Marisa
authors.initials: M
authors.lastname: Labanca
authors.affiliation: Process Biochemistry, Technical Development, Biogen, Research Triangle Park, NC, United States.
authors.forename: Lynn
authors.initials: L
authors.lastname: Conley
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D002845:Chromatography; D003545:Cysteine / Q000737:chemistry; D008956:Models, Chemical; D010084:Oxidation-Reduction; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000737:chemistry; D013205:Staphylococcal Protein A / Q000737:chemistry*; D013440:Sulfides / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D013205:Staphylococcal Protein A; D013440:Sulfides; D003545:Cysteine
keywords: Cysteine wash; Model; Monoclonal antibodies; Product heterogeneity; Protein A chromatography; Trisulfides