title: Prediction of Protein Aggregation in High Concentration Protein Solutions Utilizing Protein-Protein Interactions Determined by Low Volume Static Light Scattering.
abstract: The development of highly concentrated protein formulations is more demanding than for conventional concentrations due to an elevated protein aggregation tendency. Predictive protein-protein interaction parameters, such as the second virial coefficient B22 or the interaction parameter kD, have already been used to predict aggregation tendency and optimize protein formulations. However, these parameters can only be determined in diluted solutions, up to 20 mg/mL. And their validity at high concentrations is currently controversially discussed. This work presents a μ-scale screening approach which has been adapted to early industrial project needs. The procedure is based on static light scattering to directly determine protein-protein interactions at concentrations up to 100 mg/mL. Three different therapeutic molecules were formulated, varying in pH, salt content, and addition of excipients (e.g., sugars, amino acids, polysorbates, or other macromolecules). Validity of the predicted aggregation tendency was confirmed by stability data of selected formulations. Based on the results obtained, the new prediction method is a promising screening tool for fast and easy formulation development of highly concentrated protein solutions, consuming only microliter of sample volumes.
authors.affiliation: Department of Pharmaceutics, Friedrich-Alexander University Erlangen-Nuremberg, Freeze Drying Focus Group, Cauerstrasse 4, Erlangen 91058, Germany; Merck KGaA, Chemical and Pharmaceutical Development, Frankfurter Strasse 250, Darmstadt 64293, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Hofmann
authors.affiliation: Merck KGaA, Chemical and Pharmaceutical Development, Frankfurter Strasse 250, Darmstadt 64293, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Winzer
authors.affiliation: Merck KGaA, Chemical and Pharmaceutical Development, Frankfurter Strasse 250, Darmstadt 64293, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Weber
authors.affiliation: Department of Pharmaceutics, Friedrich-Alexander University Erlangen-Nuremberg, Freeze Drying Focus Group, Cauerstrasse 4, Erlangen 91058, Germany; GILYOS GmbH, Friedrich-Bergius-Ring 15, Würzburg 97076, Germany. Electronic address: info@gilyos.com.
authors.forename: Henning
authors.initials: H
authors.lastname: Gieseler
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism*; D000067493:Dynamic Light Scattering / Q000379:methods*; D005544:Forecasting; D007074:Immunoglobulin G / Q000737:chemistry / Q000378:metabolism*; D019999:Pharmaceutical Solutions / Q000737:chemistry / Q000378:metabolism; D066329:Protein Aggregates / Q000502:physiology*; D054730:Protein Interaction Domains and Motifs / Q000502:physiology*
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D019999:Pharmaceutical Solutions; D066329:Protein Aggregates
keywords: interaction; light scattering (static); physical stability; preformulation; protein; protein aggregation; protein formulation