Pilot Study Of Detecting Advanced Glycation End Products In Tissue And Serum With Monoclonal Antibodies

Aims: In order to explore the role of advanced glycation end products (AGEs) in pathogenesis and development of diabetic complications, and to establish an ELISA method capable of measuring the very low concentration of AGEs in serum, the monoclonal antibody (McAb) against advanced glycation end products was prepared, its immunological property and value of application were analysised.Methods: Indirect ELISA was used to screen out the hybridoma which could produce antibodies against AGEs. One of antibody-producing hybridoma wells was selected for further use through two successive subclonings by limiting dilution method. These positive cell monoclones were injected into Balb/c mice for producing the ascites. Each ascites was purified by mannan binding protein (MBP), then McAbs were identified by SDS-PAGE, indirect ELISA and western blot were used to analysis the immunological property of McAbs and the epitope which would combine with McAbs. AGEs in serum of human and in aorta,renal,heart of diabetic rats were also detected by McAbs.Results: McAbs reacted with AGEs specially and combined with non-carboxymethyl lysine (non-CML). AGEs in serum of human had been detected. AGEs in aorta,renal and heart of diabetic rats had also been detected, however, there had been any AGEs detected in healthy rats.Conclusions: McAbs(non-CML)reacted with AGEs specially, and might be of value for AGEs qualitative measurement.