Melanie Hofmann1,2, Matthias Winzer2, Christian Weber2, Henning Gieseler3. 1. Division of Pharmaceutics, Friedrich-Alexander Universität Erlangen-Nürnberg, Erlangen, Germany. 2. Merck KGaA, Darmstadt, Germany. 3. GILYOS GmbH, Würzburg, Germany.
Abstract
OBJECTIVE: Polyethylene glycol (PEG)-induced protein precipitation is often used to extrapolate apparent protein solubility at specific formulation compositions. The procedure was used for several fields of application such as protein crystal growth but also protein formulation development. Nevertheless, most studies focused on applicability in protein crystal growth. In contrast, this study focuses on applicability of PEG-induced precipitation during high-concentration protein formulation development. METHODS: In this study, solubility of three different model proteins was investigated over a broad range of pH. Solubility values predicted by PEG-induced precipitation were compared to real solubility behaviour determined by either turbidity or content measurements. KEY FINDINGS: Predicted solubility by PEG-induced precipitation was confirmed for an Fc fusion protein and a monoclonal antibody. In contrast, PEG-induced precipitation failed to predict solubility of a single-domain antibody construct. Applicability of PEG-induced precipitation as indicator of protein solubility during formulation development was found to be not valid for one of three model molecules. CONCLUSIONS: Under certain conditions, PEG-induced protein precipitation is not valid for prediction of real protein solubility behaviour. The procedure should be used carefully as tool for formulation development, and the results obtained should be validated by additional investigations.
OBJECTIVE:Polyethylene glycol (PEG)-induced protein precipitation is often used to extrapolate apparent protein solubility at specific formulation compositions. The procedure was used for several fields of application such as protein crystal growth but also protein formulation development. Nevertheless, most studies focused on applicability in protein crystal growth. In contrast, this study focuses on applicability of PEG-induced precipitation during high-concentration protein formulation development. METHODS: In this study, solubility of three different model proteins was investigated over a broad range of pH. Solubility values predicted by PEG-induced precipitation were compared to real solubility behaviour determined by either turbidity or content measurements. KEY FINDINGS: Predicted solubility by PEG-induced precipitation was confirmed for an Fc fusion protein and a monoclonal antibody. In contrast, PEG-induced precipitation failed to predict solubility of a single-domain antibody construct. Applicability of PEG-induced precipitation as indicator of protein solubility during formulation development was found to be not valid for one of three model molecules. CONCLUSIONS: Under certain conditions, PEG-induced protein precipitation is not valid for prediction of real protein solubility behaviour. The procedure should be used carefully as tool for formulation development, and the results obtained should be validated by additional investigations.
Authors: Yingda Xu; Dongdong Wang; Bruce Mason; Tony Rossomando; Ning Li; Dingjiang Liu; Jason K Cheung; Wei Xu; Smita Raghava; Amit Katiyar; Christine Nowak; Tao Xiang; Diane D Dong; Joanne Sun; Alain Beck; Hongcheng Liu Journal: MAbs Date: 2018-12-17 Impact factor: 5.857