title: Customization of copolymers to optimize selectivity and yield in polymer-driven antibody purification processes.
abstract: This manuscript describes customization of copolymers to be used for polymer-driven protein purification in bioprocessing. To understand how copolymer customization can be used for fine-tuning, precipitation behavior was analyzed for five target antibodies (mAbs) and BSA as model impurity protein, at ionic strength similar to undiluted cell culture fluid. In contrast to the use of standardized homopolymers, customized copolymers, composed of 2-acrylamido-2-methylpropane sulfonic acid (AMPS) and 4-(acryloylamino)benzoic acid (ABZ), exhibited antibody precipitation yields exceeding 90%. Additionally, copolymer average molecular weight (Mw ) was varied and its influence on precipitation yield and contaminant coprecipitation was investigated. Results revealed copolymer composition as the major driving force for precipitation selectivity, which was also dependent on protein hydrophobicity. By adjusting ABZ content and Mw of the precipitant for each of the mAbs, conditions were found that allowed for high precipitation yield and selectivity. These findings may open up new avenues for using polymers in antibody purification processes.
authors.affiliation: Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, 64287, Darmstadt, Germany; Merck KGaA, PTD, Frankfurter Strasse 250, 64293, Darmstadt, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Capito
authors.forename: Romas
authors.initials: R
authors.lastname: Skudas
authors.forename: Bernd
authors.initials: B
authors.lastname: Stanislawski
authors.forename: Harald
authors.initials: H
authors.lastname: Kolmar
mesh_terms: D010129:4-Aminobenzoic Acid / Q000737:chemistry*; D000178:Acrylamides / Q000737:chemistry*; D000476:Alkanesulfonates / Q000737:chemistry*; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification*; D018929:Cell Culture Techniques; D011232:Chemical Precipitation; D006801:Humans; D009994:Osmolar Concentration; D011108:Polymers / Q000737:chemistry*
chemical_list: D000178:Acrylamides; D000476:Alkanesulfonates; D000911:Antibodies, Monoclonal; D011108:Polymers; C056288:2-acrylamido-2-methylpropanesulfonate; D010129:4-Aminobenzoic Acid
keywords: antibody precipitation; copolymer; downstream processing; selectivity