title: Moving through three-dimensional phase diagrams of monoclonal antibodies.
abstract: Protein phase behavior characterization is a multivariate problem due to the high amount of influencing parameters and the diversity of the proteins. Single influences on the protein are not understood and fundamental knowledge remains to be obtained. For this purpose, a systematic screening method was developed to characterize the influence of fluid phase conditions on the phase behavior of proteins in three-dimensional phase diagrams. This approach was applied to three monoclonal antibodies to investigate influences of pH, protein and salt concentrations, with five different salts being tested. Although differences exist between the antibodies, this extensive study confirmed the general applicability of the Hofmeister series over the broad parameter range analyzed. The influence of the different salts on the aggregation (crystallization and precipitation) probability was described qualitatively using this Hofmeister series, with a differentiation between crystallization and precipitation being impossible, however.
authors.affiliation: Biomolecular Separation Engineering, Institute of Engineering in Life Sciences, Karlsruhe Institute of Technology, Engler-Bunte-Ring 1, 76131, Karlsruhe, Germany.
authors.forename: Natalie
authors.initials: N
authors.lastname: Rakel
authors.forename: Miriam
authors.initials: M
authors.lastname: Baum
authors.forename: Jürgen
authors.initials: J
authors.lastname: Hubbuch
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D001709:Biotechnology / Q000379:methods*; D003460:Crystallization; D057166:High-Throughput Screening Assays; D006863:Hydrogen-Ion Concentration; D007477:Ions / Q000494:pharmacology; D066329:Protein Aggregates / Q000187:drug effects; D012107:Research Design; D012965:Sodium Chloride / Q000737:chemistry / Q000494:pharmacology; D013431:Sulfates / Q000737:chemistry / Q000494:pharmacology
chemical_list: D000911:Antibodies, Monoclonal; D007477:Ions; D066329:Protein Aggregates; D013431:Sulfates; D012965:Sodium Chloride
keywords: high-throughput screening; protein phase behavior, phase diagram, monoclonal antibody, crystallization