TY - JOUR

T1 - Chemometrics in Protein Formulation

T2 - Stability Governed by Repulsion and Protein Unfolding

AU - Kulakova, Alina

AU - Augustijn, Dillen

AU - El Bialy, Inas

AU - Gentiluomo, Lorenzo

AU - Greco, Maria Laura

AU - Hervø-hansen, Stefan

AU - Indrakumar, Sowmya

AU - Mahapatra, Sujata

AU - Martinez Morales, Marcello

AU - Pohl, Christin

AU - Polimeni, Marco

AU - Roche, Aisling

AU - Svilenov, Hristo L.

AU - Tosstorff, Andreas

AU - Zalar, Matja

AU - Curtis, Robin

AU - Derrick, Jeremy P.

AU - Frieß, Wolfgang

AU - Golovanov, Alexander P.

AU - Lund, Mikael

AU - Nørgaard, Allan

AU - Khan, Tarik A.

AU - Peters, Günther H. J.

AU - Pluen, Alain

AU - Roessner, Dierk

AU - Streicher, Werner W.

AU - Van Der Walle, Christopher F.

AU - Warwicker, Jim

AU - Uddin, Shahid

AU - Winter, Gerhard

AU - Bukrinski, Jens Thostrup

AU - Rinnan, Åsmund

AU - Harris, Pernille

PY - 2023

Y1 - 2023

N2 - Therapeutic proteins can be challenging to develop due to their complexity and the requirement of an acceptable formulation to ensure patient safety and efficacy. To date, there is no universal formulation development strategy that can identify optimal formulation conditions for all types of proteins in a fast and reliable manner. In this work, high-throughput characterization, employing a toolbox of five techniques, was performed on 14 structurally different proteins formulated in 6 different buffer conditions and in the presence of 4 different excipients. Multivariate data analysis and chemometrics were used to analyze the data in an unbiased way. First, observed changes in stability were primarily determined by the individual protein. Second, pH and ionic strength are the two most important factors determining the physical stability of proteins, where there exists a significant statistical interaction between protein and pH/ionic strength. Additionally, we developed prediction methods by partial least-squares regression. Colloidal stability indicators are important for prediction of real-time stability, while conformational stability indicators are important for prediction of stability under accelerated stress conditions at 40 °C. In order to predict real-time storage stability, protein–protein repulsion and the initial monomer fraction are the most important properties to monitor.

AB - Therapeutic proteins can be challenging to develop due to their complexity and the requirement of an acceptable formulation to ensure patient safety and efficacy. To date, there is no universal formulation development strategy that can identify optimal formulation conditions for all types of proteins in a fast and reliable manner. In this work, high-throughput characterization, employing a toolbox of five techniques, was performed on 14 structurally different proteins formulated in 6 different buffer conditions and in the presence of 4 different excipients. Multivariate data analysis and chemometrics were used to analyze the data in an unbiased way. First, observed changes in stability were primarily determined by the individual protein. Second, pH and ionic strength are the two most important factors determining the physical stability of proteins, where there exists a significant statistical interaction between protein and pH/ionic strength. Additionally, we developed prediction methods by partial least-squares regression. Colloidal stability indicators are important for prediction of real-time stability, while conformational stability indicators are important for prediction of stability under accelerated stress conditions at 40 °C. In order to predict real-time storage stability, protein–protein repulsion and the initial monomer fraction are the most important properties to monitor.

U2 - 10.1021/acs.molpharmaceut.3c00013

DO - 10.1021/acs.molpharmaceut.3c00013

M3 - Journal article

C2 - 37146162

VL - 20

SP - 2951

EP - 2965

JO - Molecular Pharmaceutics

JF - Molecular Pharmaceutics

SN - 1543-8384

IS - 6

ER -