Immunochemical detection of 3-deoxyglucosone in serum.
3-Deoxyglucosone (3-DG) is a metabolite of glucose that is thought to lead to the production of advanced glycation end products in diabetes. The previous assay for 3-DG in serum was based on a multi-step protocol, including derivatization, extraction, HPLC separation, and detection. In the current studies, we established a monoclonal antibody that recognizes the 3-DG-derivative, which is generated by the reaction of 3-DG and a 2,3-diamino-benzene derivative. Attachment of a biotin moiety to the 2,3-diamino-benzene ring via a linker allowed development of a highly sensitive chemiluminescent enzyme immunoassay for 3-DG equivalents. Unlike the previous assay, this method does not require extraction of 3-DG derivatives from serum. Treatment of 3-DG in serum with the DAB-link-biotin produced a quinoxaline derivative, which was specifically recognized by the monoclonal antibody. Using this assay, we found that serum 3-DG was higher in streptozotocin-induced diabetic rats than in normal control rats (25+/-5.6 vs. 9.8+/-1.1 microg/L). This simple assay may allow the monitoring of conditions leading to the accumulation of advanced glycation end products and evaluation of the risk of complications in diabetic patients.
Uchida
Yoshiaki
Y
Fujirebio Inc., Research and Development Division, 51 Komiya-cho, Hachioji, Tokyo 192-0031, Japan. yk-uchida@fujirebio.co.jp
Kurano
Yoshihiro
Y
Endo
Tomohiro
T
Aoyama
Misao
M
Ito
Satoru
S
D000818:Animals; D015415:Biomarkers / Q000097:blood; D001774:Blood Chemical Analysis / Q000379:methods*; D003847:Deoxyglucose / Q000031:analogs & derivatives* / Q000097:blood*; D003921:Diabetes Mellitus, Experimental / Q000097:blood* / Q000139:chemically induced / Q000175:diagnosis; D017127:Glycation End Products, Advanced / Q000097:blood*; D007124:Immunoenzyme Techniques / Q000379:methods*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013311:Streptozocin
D015415:Biomarkers; D017127:Glycation End Products, Advanced; D013311:Streptozocin; D003847:Deoxyglucose; C016350:3-deoxyglucosone