title: Stabilization of a hydrophobic recombinant cytokine by human serum albumin.
abstract: 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.
authors.affiliation: Department of Pharmacy, Pharmaceutical Technology and Biopharmaceutics, Ludwig-Maximilians-University Munich, Germany. Andrea.Hawe@cup.uni-muenchen.de
authors.forename: Andrea
authors.initials: A
authors.lastname: Hawe
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Friess
mesh_terms: D011232:Chemical Precipitation; D016207:Cytokines / Q000737:chemistry*; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D008027:Light; D009391:Nephelometry and Turbidimetry; D009994:Osmolar Concentration; D010316:Particle Size; D011994:Recombinant Proteins / Q000737:chemistry; D012542:Scattering, Radiation; D012709:Serum Albumin / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry; D013050:Spectrometry, Fluorescence