Prediction and Reduction of the Aggregation of Monoclonal Antibodies.
Protein aggregation remains a major area of focus in the production of monoclonal antibodies. Improving the intrinsic properties of antibodies can improve manufacturability, attrition rates, safety, formulation, titers, immunogenicity, and solubility. Here, we explore the potential of predicting and reducing the aggregation propensity of monoclonal antibodies, based on the identification of aggregation-prone regions and their contribution to the thermodynamic stability of the protein. Although aggregation-prone regions are thought to occur in the antigen binding region to drive hydrophobic binding with antigen, we were able to rationally design variants that display a marked decrease in aggregation propensity while retaining antigen binding through the introduction of artificial aggregation gatekeeper residues. The reduction in aggregation propensity was accompanied by an increase in expression titer, showing that reducing protein aggregation is beneficial throughout the development process. The data presented show that this approach can significantly reduce liabilities in novel therapeutic antibodies and proteins, leading to a more efficient path to clinical studies.
VIB Switch Laboratory, Herestraat 49, B-3000 Leuven, Belgium; Department of Cellular and Molecular Medicine, KU Leuven, Herestraat 49, PO 802, B-3000 Leuven, Belgium.
Rob
R
van der Kant
Boehringer Ingelheim Pharma GmbH & Co. KG, 88400, Biberach/Riss, Germany.
Anne R
AR
Karow-Zwick
VIB Switch Laboratory, Herestraat 49, B-3000 Leuven, Belgium; Department of Cellular and Molecular Medicine, KU Leuven, Herestraat 49, PO 802, B-3000 Leuven, Belgium.
Joost
J
Van Durme
Boehringer Ingelheim Pharma GmbH & Co. KG, 88400, Biberach/Riss, Germany.
Michaela
M
Blech
VIB Switch Laboratory, Herestraat 49, B-3000 Leuven, Belgium; Department of Cellular and Molecular Medicine, KU Leuven, Herestraat 49, PO 802, B-3000 Leuven, Belgium.
Rodrigo
R
Gallardo
Boehringer Ingelheim Pharma GmbH & Co. KG, 88400, Biberach/Riss, Germany.
Daniel
D
Seeliger
Boehringer Ingelheim Pharma GmbH & Co. KG, 88400, Biberach/Riss, Germany.
Kerstin
K
Aßfalg
EM-platform VIB Bio Imaging Core, VIB-KU Leuven, Herestraat 49, B-3000 Leuven.
Pieter
P
Baatsen
Department of Pharmaceutical and Pharmacological Sciences, Laboratory for Therapeutic and Diagnostic Antibodies, KU Leuven, Herestraat 49, PO 820, B-3000 Leuven, Belgium.
Griet
G
Compernolle
Department of Pharmaceutical and Pharmacological Sciences, Laboratory for Therapeutic and Diagnostic Antibodies, KU Leuven, Herestraat 49, PO 820, B-3000 Leuven, Belgium.
Ann
A
Gils
Boehringer Ingelheim Pharma GmbH & Co. KG, 88400, Biberach/Riss, Germany.
Joey M
JM
Studts
Boehringer Ingelheim Pharma GmbH & Co. KG, 88400, Biberach/Riss, Germany.
Patrick
P
Schulz
Boehringer Ingelheim Pharma GmbH & Co. KG, 88400, Biberach/Riss, Germany.
Patrick
P
Garidel
VIB Switch Laboratory, Herestraat 49, B-3000 Leuven, Belgium; Department of Cellular and Molecular Medicine, KU Leuven, Herestraat 49, PO 802, B-3000 Leuven, Belgium. Electronic address: joost.schymkowitz@switch.vib-kuleuven.be.
Joost
J
Schymkowitz
VIB Switch Laboratory, Herestraat 49, B-3000 Leuven, Belgium; Department of Cellular and Molecular Medicine, KU Leuven, Herestraat 49, PO 802, B-3000 Leuven, Belgium. Electronic address: frederic.rousseau@switch.vib-kuleuven.be.
Frederic
F
Rousseau
D000465:Algorithms; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016466:CHO Cells; D019295:Computational Biology / Q000379:methods*; D003198:Computer Simulation; D003412:Cricetulus; D006801:Humans; D009154:Mutation; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods; D013329:Structure-Activity Relationship
D000911:Antibodies, Monoclonal
monoclonal antibody; protein aggregation; protein engineering; protein folding