Development of a Rapid Assay for Advanced Glycosylation End Products in Human Serum

• Main Authors: Chwei-Yuh Lin, 林垂裕
• Other Authors: Horng-Mo Lee
• Format: Others
• Language: zh-TW
• Published: 2001
• Online Access: http://ndltd.ncl.edu.tw/handle/28043674942586263907

碩士 === 台北醫學院 === 生物醫學技術研究所 === 89 === Non-enzymatic reaction of protein and carbohydrate may produce a series of brown fluorescent advanced glycosylation end products(AGEs), AGEs accumulation in tissue have been linked to diabetic complications. However, convenient and rapid assay for serum LMW-AGE is currently unavailable. In the present study, we produced RNase-AGEs by incubating RNase with high concentration of glucose, The RNase-derived AGEs were then used to immunize rabbit and produce AGEs-specific polyclonal antibodies. We used the anti-AGEs antibodies to coated latex and used it to develop an immunoturbidimetry, which can be use in autoanalyzer(eg. Olympus, AU-600). By the calibration curve(R2 =0.9953), recovery rate(98.4 %), The precision was better than those in competitive ELISA, with a within day of C.V=2.9 %; between day of C.V=6.1 %. The interference was verified by hemolysis, icterus, lipemic serum(cholesterol <320 mg/dl; triglyceride<500mg/dl) not interfered respectively. We are currently using this assay to measure serum LMW-AGEs in normal individuals(young 4.6±1.5, n=39; elderly 4.9±1.4, n=40), diabetic patients(7.8±4.8, n=89) and patients with end stage renal disease without(17.2±7.7, n=28) or with DM(16.3±6.0, n=12). The assay will be proven useful in predict the diabetic complications in diabetes.