Slot dot-blot as an effective method for detection of AGEs present in sera in trace amounts

AIM: The aim of studies was to develop immunochemical methods for the detection of proteins glycated by glucose (AGE1), methylglyoxal (AGE3), glycolaldehyde (AGE4) with using the following antibodies: anti-AGE1, anti-AGE3 and anti-AGE4 (Cosmo Bio, Japan). According to our knowledge, any work with using these antibodies has been published yet. First, we used ELISA, however, it seemed to be insufficiently sensitive. We have developed conditions of the method of the slot dot-blot, which uses larger amounts of serum. However, beneficially the slot dot-blot allows also for semi-quantitative determination of AGE1. METHODS: In order to determine the presence of AGE1, AGE3 and AGE4 in sera of diabetic patients we used the slot dot-blot method. As the secondary antibodies we used goat anti-mouse immunoglobulins coupled with HRP, no cross-reactive with human IgG. Additionally we carried out the slot dot-blot of sera against secondary antibodies, as a control blot. RESULTS: AGE1 level in serum of diabetic patients was significantly higher than AGE3 and AGE4. Slot dot-blot was effective method for detection of AGE4, present in sera in trace amounts. CONCLUSIONS: The slot dot-blot method is an effective method for detection of AGE1, AGE3 and AGE4 in blood serum