title: Effects of sugar additives on protein stability of recombinant human serum albumin during lyophilization and storage.
abstract: Few researches on the protein stabilization of recombinant human serum albumin (rHSA) have been done. In the present study, we assessed the impact of sugar lyoprotectants on the protein stability of lyophilized rHSA (65 KDa) in the solid state. For the assessment, rHSA was formulated with sucrose and trehalose, respectively, alone or in combination with mannitol, which were lyophilized and stored at 35 degrees C. Degradation and aggregation of the resulting lyophilized formulations was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Induction of amorphous state by the lyophilactants with rHSA was determined by differential scanning calorimetry (DSC). The protein secondary structure of the rHSA in the formulations was analyzed by Fourier transform infrared spectroscopy (FT-IR). Results from SDS-PAGE analysis displayed that mannitol formulation caused aggregation resulting in a few bands that were greater than 65 KDa, whereas sucrose and trehalose formulations revealed no such aggregation. However, the aggregation of the protein decreased when mannitol was combined with sucrose or trehalose. DSC measurement supported the electrophoresis data showing that sucrose and trehalose formed complete amorphous state, but mannitol induced a partial amorphous state. These data indicate during lyophilization the most effective protein protection against aggregation was provided by sucrose and trehalose. The protection lasted during 4 months storage at 35 degrees C. FT-IR analysis displayed that the sucrose formulation inhibited deamidation. In conclusion, our data suggest that sucrose and trehalose as additives seems to be sufficient to protect from lyophilization of rHSA protein and also maintain its stability in the solid state during storage.
authors.affiliation: Department of ImmunoMicrobiology, College of Pharmacy, Dongduk Women's University, Seoul 136-714, Korea. ymhan@dongduk.ac.kr
authors.forename: Yongmoon
authors.initials: Y
authors.lastname: Han
authors.forename: Byung-Seok
authors.initials: BS
authors.lastname: Jin
authors.forename: Sang-Bong
authors.initials: SB
authors.lastname: Lee
authors.forename: Yeowon
authors.initials: Y
authors.lastname: Sohn
authors.forename: Jee-Won
authors.initials: JW
authors.lastname: Joung
authors.forename: Jue-Hee
authors.initials: JH
authors.lastname: Lee
mesh_terms: D002152:Calorimetry, Differential Scanning; D002241:Carbohydrates / Q000494:pharmacology*; D003460:Crystallization; D004355:Drug Stability; D004356:Drug Storage; D005612:Freeze Drying; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry*; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D002241:Carbohydrates; D011994:Recombinant Proteins; D012709:Serum Albumin