Stabilization of a hydrophobic recombinant cytokine by human serum albumin

The objective was to evaluate the impact of pH and NaCl content on aggregation, particle formation, and solubility of a hydrophobic recombinant human cytokine in formulations with human serum albumin (HSA) as stabilizing excipient. While cytokine-HSA formulations were stable at physiological pH, a tremendous increase in turbidity at pH 5.0, close to the isoelectric point of HSA was caused by a partially irreversible precipitation. By dynamic light scattering (DLS), disc centrifugation, atomic force microscopy (AFM), and light obscuration it could be shown that the turbidity was mainly caused by particles larger than 120 nm. SDS–PAGE provided evidence that the precipitation at pH 5.0 was mainly caused by the cytokine. The HSA-stabilizers Na-octanoate and Na-N-acetyltryptophante were less effective in preventing the turbidity increase of unstabilized-HSA compared to NaCl. The interactions between HSA and cytokine were weakened by NaCl, as determined by fluorescence spectroscopy. The positive effect of NaCl on the formulation could be attributed to a direct stabilization of HSA and weaker interactions between HSA and the cytokine, which in consequence provided an overall stabilization of the cytokine. © 2007 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 96: 2987–2999, 2007