Preparation of insulin monoclone antibody and its application in insulin assay in patients with type 2 diabetes mellitus / 国际检验医学杂志

ABSTRACT

Objective The monoclone antibody against insulin was prepared and used to measure plasma level of insulin in patients with type 2 diabetes mellitus. Methods Balb/c mice were immunedwith cross-linking insulin-BSA as immunogen. Four monoelone cell strains stably secreting insulin monoclone antibody were prepared by applying hybridoma technique. The valence, subclass and affini-ty constant of the monoclone antibody were analyzed. The serum insulin content was measured with competitive inhibition ELISA in 90 patients with type 2 diabetes mellitus and 90 healthy controls. Re-suits The affinity constant of the established monoclone antibody reached to 1011. The subclass of 1 strain was Igab(k),and the other 3 strains were Ig1(k) srbclass.The mean level of insulin was significantly lower in patients with type 2 diabetes mellitus than that of healthy controls. Conclusion These results implicated that the insulin insufficient might be due to the depletion of the pancreatic be-ta cell function of patients with type 2 diabetes mellitus, and the pathogenetic condition might be alle-viated by supplying insulin. About 10% patients are with higher insulin level than healthy controls,which may be due to insulin resistance. It is inadvisable to treat these patients with insulin therapy.