Effect of polyols on polyethylene glycol (PEG)-induced precipitation of proteins: Impact on solubility, stability and conformation.
Effect of polyols on the solubility of bovine serum albumin (BSA) in the presence of polyethylene glycols (PEGs) was investigated in order to strengthen the understanding of the observed effects of polyols and PEGs on protein properties in solution. Effect of polyols and/or PEGs on the thermodynamic (conformational) stability of BSA was measured using DSC and circular dichroism (CD). Glucose, sucrose, raffinose, glycerol and sorbitol, all reduced the extent of protein precipitation. Solubility of BSA in the presence of ethylene glycol increased in the case of PEG 1450 and PEG 8000, but was unaffected in the case of PEG 400. DSC studies indicated that smaller PEGs have destabilizing influence on protein structure. CD studies showed that smaller PEGs (ethylene glycol) induce subtle unfolding while stabilizing polyols induce subtle compaction. Results show that, effect of polyols on the apparent solubility of the protein correlates with their effect on the thermodynamic stability of the protein, smaller PEGs are not appropriate for estimating the activity of proteins in saturated solutions, and subtle changes in protein conformation can significantly affect protein precipitation. Though smaller PEGs have weak attractive interactions with protein molecules, perturbation of protein structure by PEGs can be balanced by utilizing appropriate stabilizing solutes.
Kumar
Vineet
V
Department of Pharmaceutical Sciences, University of Connecticut, Storrs, CT 06269, USA.
Sharma
Vikas K
VK
Kalonia
Devendra S
DS
D000818:Animals; D002152:Calorimetry, Differential Scanning; D002417:Cattle; D011232:Chemical Precipitation; D002942:Circular Dichroism; D004355:Drug Stability; D008970:Molecular Weight; D011092:Polyethylene Glycols / Q000737:chemistry*; D017433:Protein Structure, Secondary / Q000187:drug effects; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D012995:Solubility; D013402:Sugar Alcohols / Q000737:chemistry*; D013816:Thermodynamics
D013402:Sugar Alcohols; D012710:Serum Albumin, Bovine; D011092:Polyethylene Glycols