Influence of liver isoferritin profile on quantitative measurement of total ferritin protein in human liver, with emphasis on iron overload.
Ferritin was isolated from normal human liver and from iron-loaded human liver by gel chromatography and by ultracentrifugation. From each of these ferritin batches several isoferritin fractions were isolated by preparative isoelectric focusing. It was our aim to have at our disposal isoferritin fractions with relatively large pI ranges but distinctly different isoferritin profiles on analytical isoelectric focusing. These fractions were compared for their immunoreactivity. The total protein in the isoferritin fractions was measured by the nitrogen content. The immunoreactivity of the isoferritin fractions was measured as the ratio of the reaction in immunoassays to the nitrogen content of these fractions. We used radial immunodiffusion and enzyme-linked immunoassay to measure immunoreactivity. The immunoreactivity did not change obviously with the isoferritin composition of the isolated fractions. It is concluded that pathological changes in the isoferritin composition that might occur in liver ferritin during iron overload does not significantly influence the quantitative measurement of liver ferritin protein by immunological methods.
D004591:Electrophoresis, Polyacrylamide Gel; D005293:Ferritins / Q000302:isolation & purification*; D006801:Humans; D007118:Immunoassay; D007501:Iron / Q000506:poisoning*; D007525:Isoelectric Focusing; D008099:Liver / Q000378:metabolism*; D013194:Staining and Labeling
D005293:Ferritins; D007501:Iron