Revisit PEG-Induced Precipitation Assay for Protein Solubility Assessment of Monoclonal Antibody Formulations

Purpose: Protein solubility is an important attribute of pharmaceutical monoclonal antibody (MAb) formulations, particularly at high MAb concentrations. PEG-induced protein precipitation has been routinely used to assess protein solubility. To provide insights for better understanding and implementation of PEG-induced protein precipitation assay, this work compares different solubility measures and examines their relevance to loss of protein solubility in concentrated formulations.

Methods: Solubility of a MAb in 15 formulations was evaluated using PEG-induced precipitation assay. Three apparent protein solubility measures, the middle-point and onset PEG concentrations (c_mid and c_onset) as well as the binding free energy (μ_B), were obtained from the PEG-induced protein precipitation assay and compared to the DLS protein interaction parameter (k_D). Visual inspection of loss of protein solubility in concentrated formulations during storage was used to further examine the discrepancy of protein solubility ranking by these measures.

Results: PEG-induced precipitation assay predicted overall protein solubility ranking similar to that by DLS k_D. However, for three formulations with ionic excipients NaCl, Arg·Cl, and Arg·Glu·Cl, PEG-induced precipitation assay yielded more accurate predictions compared to DLS k_D measurements. Furthermore, μ_B showed superior ability in distinguishing protein solubility for these formulations.

Conclusions: This study demonstrated good correlations between the protein solubility measures obtained from PEG-induced precipitation experiments and DLS k_D measurement. It also provides one example in which protein solubility ranking by binding free energy is more accurate than the other measures. The results support the theoretical proposition that μ_B has a potential to serve as standard protein solubility measure.