Role of trehalose in moisture-induced aggregation of bovine serum albumin.
Moisture-induced aggregation has been identified as a key problem in the long term storage stability of therapeutic proteins. In the present work, we have investigated the impact of the disaccharide trehalose on the aggregation behavior of a model protein, bovine serum albumin (BSA) under moist conditions. About 50% aggregation of BSA was observed at a moisture level of 8 microl/10mg protein. Including trehalose in the protein sample caused a significant reduction in aggregation. We address the probable mechanisms for the protective effect of trehalose by considering the various hypotheses that have been proposed in the literature. The techniques that have been used include denaturing and non-denaturing gel electrophoresis and tryptophan intrinsic fluorescence. The nature of the aggregates was studied by carrying out electrophoresis of the aggregated protein in the presence of reducing and chaotropic reagents. The interaction studies of aggregated BSA with Thioflavin T and CongoRed indicate the possibility of amyloid type of character in the former. These studies may explain the protective role of trehalose under conditions where the storage stability of therapeutic proteins is compromised.
Jain
Nishant K
NK
Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar, Punjab 160062, India.
Roy
Ipsita
I
D052160:Benzothiazoles; D003224:Congo Red; D004355:Drug Stability; D004356:Drug Storage; D004591:Electrophoresis, Polyacrylamide Gel; D005079:Excipients; D005456:Fluorescent Dyes; D006813:Humidity; D011489:Protein Denaturation; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013844:Thiazoles; D014199:Trehalose / Q000737:chemistry*; D014364:Tryptophan / Q000737:chemistry
D052160:Benzothiazoles; D005079:Excipients; D005456:Fluorescent Dyes; D013844:Thiazoles; C009462:thioflavin T; D012710:Serum Albumin, Bovine; D003224:Congo Red; D014364:Tryptophan; D014199:Trehalose