Trehalose and hyaluronic acid coordinately stabilized freeze-dried pancreatic kininogenase.
The ability and mechanisms of stabilization of freeze-dried formulations of pancreatic kininogenase (PKase) by carbohydrates were evaluated. Activity and structure of PKase were examined after freeze-drying and rehydration in presence with or without a carbohydrate. Addition of trehalose, lactose, sucrose, hyaluronic acid (HA) or a combination of trehalose and HA to PKase formulations prior to freeze-drying step increases the stability of PKase during freeze-drying, storage and rehydration as measured by activity preservation. The combination of trehalose and HA is the most effective for the stabilization of PKase. Addition of HA alone to a formulation does not affect protein structure, but it increases glass-transition temperature (Tg) and stability of lyophilized PKase in presence of trehalose during dehydration, storage and rehydration processes. Therefore, trehalose and HA offer complementary properties that improve the stability of PKase during dehydration, storage and rehydration.
Zhang
Yuhua
Y
College of Food Science & Nutritional Engineering, China Agricultural University, Beijing, China.
Ji
Baoping
B
Ling
Peixue
P
Zhang
Tianmin
T
D002152:Calorimetry, Differential Scanning; D002626:Chemistry, Pharmaceutical; D002942:Circular Dichroism; D004356:Drug Storage; D004795:Enzyme Stability; D005079:Excipients / Q000737:chemistry*; D005612:Freeze Drying; D006820:Hyaluronic Acid / Q000737:chemistry*; D006860:Hydrogen Bonding; D007610:Kallikreins / Q000737:chemistry* / Q000378:metabolism; D007785:Lactose / Q000737:chemistry; D010179:Pancreas / Q000201:enzymology*; D017510:Protein Folding; D017433:Protein Structure, Secondary; D012995:Solubility; D017550:Spectroscopy, Fourier Transform Infrared; D013395:Sucrose / Q000737:chemistry; D013997:Time Factors; D044366:Transition Temperature; D014199:Trehalose / Q000737:chemistry*
D005079:Excipients; D013395:Sucrose; D006820:Hyaluronic Acid; D014199:Trehalose; D007610:Kallikreins; D007785:Lactose