Figure 9
Evaluation of specificity and stability of the ECL-based assay for human IA. A) Samples from five IA+ and one IA- donors as determined by RBA were incubated with an increasing percentage of non-biotinylated insulin prior to addition of biotinylated insulin. B) Example dataset from the competition assay for one IA+ and one IA- donor. IA value for each donor was determined from the mean signal from triplicate wells with insulin divided by the mean signal from triplicate background wells without insulin. C) Curve generated by ROC analysis of the ability of the ECL assay to distinguish between samples of RBA determined IA status. Analysis was performed using Prism software. D) Samples from IA+ and IA- donors were used to evaluate the reproducibility of the ECL assay across multiple batches of reagents. TAG-IgG = ruthenium labeled goat anti-human IgG secondary antibody. HAD = Human Assay Diluent.