Abstract
Enzymeimmunoassay, using monoclonal antibody coated polystyrene bead and insulin-peroxidase conjugate, to measure serum insulin levels was developed. A murine monoclonal antibody was prepared for the antibody coated onto bead and insulin-peroxidase conjugate was prepared by periodate method. The optimum molar ratio of insulin: peroxidase was 1.5: 1 for the conjugation reaction for high yield and good assay performances.
Intra-and inter-assay coefficient of variation (CV) were 2.3-13.6% and 3.4-14.4%, respectively.The average recovery using five serum samples were 97.5±5.5% and the dilution study showed good linearity. The assay was satisfactorily correlated with a commercially available radioimmunoassay kit (r=0.98). From the precision profiles in comparison with four commercial insulin enzymeimmunoassay kits, present assay showed the best precision of all kits. It was considered that present assay system could be utilized for a routine assay for serum insulin.
