Separation of Freezing- and Drying-Induced Denaturation of Lyophilized Proteins Using Stress-Specific Stabilization: II. Structural Studies Using Infrared Spectroscopy

摘要: The conformation of two labile enzymes, lactate dehydrogenase and phosphofructokinase, has been examined in the aqueous lyophilized states, using infrared spectroscopy. In preceding paper it was demonstrated that a stress-specific stabilization scheme, which employs combination cryoprotectant (polyethylene glycol) compound protects dried protein (sugars or mannitol), can be used to optimize recovery activity these enzymes upon freeze-drying rehydration. purpose present study is determine effects additives on during lyophilization. Lyophilization absence stabilizers observed induce significant conformational changes both enzymes. Addition 10 mM mannitol, lactose, trehalose 1% polyethylene glycol enzyme solutions attenuated unfolding, but spectral differences for state are still when compared conformation. any one does not improve activity. However, PEG either added, native structure preserved lyophilization essentially full enzymatic recovered reconstitution. ability preserve correlates directly with It appears proteins, preservation requisite following This demonstrates spectroscopic technique rapid useful method studying aid determining optimal conditions proteins