Abstract
A quantitative method to measure islet cell surface antibodies (ICSA) in human patients, using the enzyme-linked immunosorbent assay (ELISA), has been developed. Heat-inactivated human sera were incubated with monolayer cultures of a human pancreatic B cell clone (JHPI-1, 104 cells/well) for 60 min at 37°C. The cells were then washed and incubated with peroxidaseconjugated anti-human IgG rabbit serum for 30 min at room temperature. After incubation, the optical density was measured with a microplate calorimeter at a wavelength of 492 nm. The intraassay and interassay coefficients of variation were 6.6% and 13.0%, respectively. There was a significant relationship between the results of ELISA and immunofluorescence assay (IF)(p<0.005). With this method, ICSA were detected in 34.7% of insulin-dependent diabetes mellitus patients. We developed a useful method for measurement of ICSA quantitatively.
