Tungsten-induced protein aggregation: solution behavior

J Pharm Sci. 2009 Dec;98(12):4695-710. doi: 10.1002/jps.21778.


Tungsten has been associated with protein aggregation in prefilled syringes (PFSs). This study probed the relationship between PFSs, tungsten, visible particles, and protein aggregates. Experiments were carried out spiking solutions of two different model proteins with tungsten species obtained from the extraction of tungsten pins typically used in syringe manufacturing processes. These results were compared to those obtained with various soluble tungsten species from commercial sources. Although visible protein particles and aggregates were induced by tungsten from both sources, the extract from tungsten pins was more effective at inducing the formation of the soluble protein aggregates than the tungsten from other sources. Furthermore, our studies showed that the effect of tungsten on protein aggregation is dependent on the pH of the buffer used, the tungsten species, and the tungsten concentration present. The lower pH and increased tungsten concentration induced more protein aggregation. The protein molecules in the tungsten-induced aggregates had mostly nativelike structure, and aggregation was at least partly reversible. The aggregation was dependent on tungsten and protein concentration, and the ratio of these two and appears to arise through electrostatic interaction between protein and tungsten molecules. The level of tungsten required from the various sources was different, but in all cases it was at least an order of magnitude greater than the typical soluble tungsten levels measured in commercial PFS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Buffers
  • Chromatography, Gel
  • Circular Dichroism
  • Hydrogen-Ion Concentration
  • Light
  • Mass Spectrometry
  • Particle Size
  • Protein Conformation
  • Proteins / chemistry*
  • Scattering, Radiation
  • Solutions
  • Spectrophotometry, Infrared
  • Spectrophotometry, Ultraviolet
  • Spectrum Analysis, Raman
  • Tungsten / chemistry*


  • Buffers
  • Proteins
  • Solutions
  • Tungsten