Development of an automated immunoassay for advanced glycosylation end products in human serum.
OBJECTIVE
Nonenzymatic reaction of protein and carbohydrate produce a series of brown fluorescent advanced glycosylation end products (AGEs). However, a convenient and rapid assay for serum AGEs level is currently unavailable.


METHODS
We raised AGEs-specific polyclonal antibodies, which were used to develop a fully automated, noncompetitive, homogeneous, light-scattering immunoassay for serum AGEs.


RESULTS
The assay requires a sample volume of 2 microL and takes a reaction time of 2 min. The coefficient of variation range from 1.8 to 6.1%, and the mean recovery rate was 98.6%. Comparative analysis shows moderate correlation with competitive ELISA (r = 0.8209, n = 52). The mean +/- SD concentration of AGEs in young and in older healthy subjects were 4.6 +/- 1.5 (n = 39) and 4.9 +/- 1.4 (n = 40), respectively. The level of AGEs was significantly higher in serum from patients with type II DM 7.8 +/- 4.8 (n = 89) than that from the normal subjects (p < 0.05).


CONCLUSIONS
The automatic immunoassay for AGEs is appropriate for clinical use.
Lin
Chuai-Yu
CY
Graduate Institute of Biomedical Technology, Taipei Medical University, Taipei, Taiwan, Republic of China.
Chen
Chuan-Sern
CS
Shieh
Ming-Song
MS
Wu
Chih-Hsiung
CH
Lee
Horng-Mo
HM
D000368:Aged; D001331:Automation / Q000379:methods; D003920:Diabetes Mellitus / Q000097:blood; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D017127:Glycation End Products, Advanced / Q000097:blood*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007124:Immunoenzyme Techniques; D007674:Kidney Diseases / Q000097:blood; D008297:Male; D008875:Middle Aged; D012680:Sensitivity and Specificity
D017127:Glycation End Products, Advanced