title: Development of a monoclonal antibody-based ELISA system for glyceraldehyde-derived advanced glycation end products.
abstract: We have previously found that glyceraldehyde-derived advanced glycation end products (glycer-AGEs) elicit oxidative stress generation and evoke inflammatory and thrombotic reactions through their higher binding affinity to RAGE (receptor for AGEs), thereby playing a role in vascular complications in diabetes. Furthermore, circulating levels of glycer-AGEs are elevated in diabetes. We characterized a monoclonal antibody (mAb) raised against glycer-AGEs and prepared its specific ELISA system in human serum. We developed here mAb reacted specifically with glycer-AGEs or glyceraldehyde-derived pyridinium, but not other structurally identified AGEs or AGE precursors. The mAb not only completely neutralized the deleterious effects of glycer-AGEs on endothelial cells, but also detected glycer-AGEs in the aorta of type 2 diabetic rats. Intra and inter-assay coefficient variations of the ELISA were 6 and 2.6%, respectively. ELISA linearity was shown intact within 5-fold dilution, and recovery ratio of added glycer-AGEs was 88-117%. Results of serum and plasma were comparable, and repeated freeze-thawing of samples did not affect the results (90.1-112.4%). Serum glycer-AGEs levels in 30 healthy subjects evaluated by the ELISA were strongly correlated with those by polyclonal Ab-based one (r=0.82). Our present study suggests the clinical utility of mAb for evaluating glycer-AGE levels in both tissue and serum.
authors: Department of Pathophysiology and Therapeutics of Diabetic Vascular Complications, Kurume University School of Medicine, Kurume, Japan.
authors: Takanori
authors: T
authors: Matsui
authors: Boditech Med Inc., Chuncheon, South Korea.
authors: Hoo Don
authors: HD
authors: Joo
authors: Boditech Med Inc., Chuncheon, South Korea.
authors: Jae Min
authors: JM
authors: Lee
authors: Boditech Med Inc., Chuncheon, South Korea.
authors: Sung Mi
authors: SM
authors: Ju
authors: Boditech Med Inc., Chuncheon, South Korea.
authors: Wang Hong
authors: WH
authors: Tao
authors: Department of Chemistry, Kurume University School of Medicine, Kurume, Japan.
authors: Yuichiro
authors: Y
authors: Higashimoto
authors: Department of Medicine, Kurume University School of Medicine, Kurume, Japan.
authors: Kei
authors: K
authors: Fukami
authors: Department of Pathophysiology and Therapeutics of Diabetic Vascular Complications, Kurume University School of Medicine, Kurume, Japan. Electronic address: shoichi@med.kurume-u.ac.jp.
authors: Sho-ichi
authors: S
authors: Yamagishi
mesh_terms: D000368:Aged; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D042783:Endothelial Cells; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D017127:Glycation End Products, Advanced / Q000097:blood / Q000276:immunology / Q000378:metabolism; D005985:Glyceraldehyde / Q000031:analogs & derivatives*; D064368:Healthy Volunteers; D006801:Humans; D008297:Male; D008875:Middle Aged
chemical_list: D000911:Antibodies, Monoclonal; D017127:Glycation End Products, Advanced; D005985:Glyceraldehyde
keywords: AGEs; Diabetes; ELISA; Endothelial cells; Monoclonal antibody