Glass Leachables as a Nucleation Factor for Free Fatty Acid Particle Formation in Biopharmaceutical Formulations.

Surfactants are essential components in protein formulations protecting them against interfacial stress. One of the current industry-wide challenges is enzymatic degradation of parenteral surfactants such as polysorbate 20 (PS20) and polysorbate 80, which leads to the accumulation of free fatty acids (FFAs) potentially forming visible particles over the drug product shelf-life. While the concentration of FFAs can be quantified, the time point of particle formation remains unpredictable. In this work, we studied the influence of glass leachables as nucleation factors for FFA particle formation. We demonstrate the feasibility of nucleation of FFA particles in the presence of inorganic salts like NaAlO and CaCl simulating relevant glass leachables. We further demonstrate FFA particle formation depending on relevant aluminum concentrations. FFA particle formation was subsequently confirmed with lauric/myristic acid in the presence of different quantities and compositions of glass leachables obtained by several sterilization cycles using different types of glass vials. We further verified the formation of particles in aged protein formulation containing degraded PS20 through the spiking of glass leachables. Particles were characterized as a complex of glass leachables, such as aluminum and FFAs. Based on our findings, we propose a likely pathway for FFA particle formation that considers specific nucleation factors.