Thermal stability of low molecular weight urokinase during heat treatment. II. Effect of polymeric additives

Pharm Res. 1994 Jul;11(7):1004-8. doi: 10.1023/a:1018935420680.

Abstract

Turbidimetric or light scattering assays can be used to determine the extent of aggregation in protein formulations. Using low molecular weight urokinase (LMW-UK) as a model protein, the effect of polymeric additives on heat-induced aggregation was evaluated. Previous work has shown that under 60 degrees C heat treatment, LMW-UK initially denatures and the unfolded protein associates to form soluble aggregates. Eventually, these aggregates associate to form a precipitate. The effects of polymers on the initial aggregation phase was examined. Hydroxyethyl (heta) starch, polyethylene glycol 4000, and gelatin were found to be effective, concentration-dependent inhibitors of aggregation, whereas polyvinylpyrrolidone (PVP) and polyethylene glycol 300 were ineffective. Overall, the effect of polymeric additives on the stability of thermally-stressed LMW-UK can be accounted for by preferential exclusion of the solute from the surface of the protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Enzyme Stability
  • Glycerol / pharmacology
  • Hot Temperature
  • Light
  • Molecular Weight
  • Nephelometry and Turbidimetry
  • Polyethylene Glycols
  • Polymers*
  • Povidone
  • Protein Denaturation
  • Protein Folding
  • Scattering, Radiation
  • Starch / pharmacology
  • Urokinase-Type Plasminogen Activator / chemistry*
  • Urokinase-Type Plasminogen Activator / drug effects

Substances

  • Polymers
  • Polyethylene Glycols
  • Starch
  • Urokinase-Type Plasminogen Activator
  • Povidone
  • Glycerol