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Abstract

The present invention relates to methods for the diagnosis and monitoring of diseases and disorders associate with advanced glycosylation endproducts (AGE) formation, such as diabetes and the ageing process. In particular, the invention is directed to detecting AGE-modified hemoglobin (Hb-AGE) for the foregoing purposes, and in improved assay therefore. The method involves diluting the sample in a dilution buffer, which dilution buffer comprises an anionic protein denaturing detergent at a concentration sufficient to denature hemoglobin-AGE without interfering in binding of reagents with hemoglobin-AGE the dilution buffer may also include a non-ionic surfactant at a concentration sufficient to facilitate detection of hemoglobin-AGE; and a denaturing agent at a concentration sufficient to denature hemoglobin-AGE and increase assay sensitivity, without denaturing binding of reagents to hemoglobin-AGE. After diluting the sample in the dilution buffer, the sample is contacted with means for detecting the presence of hemoglobin-AGE in the sample, and the presence of hemoglobin-AGE in the sample is detected with the detection means. Dilution buffers and kits for practicing the invention are also provided. In specific examples, the level of AGE in hemoglobin in samples from human and rat normal subjects and diabetic subjects is detected. The results obtained from human samples show a high degree of correlation between the level of hemoglobin-AGE in a sample and the level of hemoglobin-A.sub.1c in a sample. Most importantly, the invention is used to detect the 'aminoguanidine effect,' which is the decrease in the level of hemoglobin-AGE in a sample from a subject undergoing therapy with the AGE-inhibitor aminoguanidine.

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First Claim