Stability of radioiodinated ferritin.

Ferritin which had been radioiodinated using chloramine T exhibited marked instability on storage at 4 degrees C. Both [125I]human liver and heart ferritins showed a similar rate of decline in immunoreactivity (t 1/2 = 20-23 days) indicating that deterioration with storage was not a function of isoferritin composition. The decrease in radioactivity associated with ferritin was due not only to loss of 125I from the molecule but also to protein degradation as shown by enzyme-linked immunoassay and gel filtration. The degradation products had an Mr of at least 69,000 although low Mr material could be identified by gel filtration when a marked reduction in immunoreactivity had occurred. Ferritin instability was much more pronounced than when other proteins such as immunoglobulin and albumin were radioiodinated with chloramine T. These observations indicate that when performing in vivo and in vitro studies with labeled ferritin, degradation of the protein during storage should be carefully monitored and the protein repurified before use.