title: Detection of chromogranin A in the adrenal gland extracts of different animal species by an enzyme-linked immunosorbent assay using Thomsen-Friedenreich antigen-specific Amaranthus caudatus lectin.
abstract: The reactivity of different lectins with crude chromogranin A (CgA) obtained from different animals, namely, cow, horse, dog, pig, and dolphin, was examined to identify lectin(s) that would be useful as coating reagent(s) in a sandwich enzyme-linked immunosorbent assay (ELISA). Of the different lectins studied, the Amaranthus caudatus lectin (ACA), which is specific for the Thomsen-Friedenreich (T)-antigen (Galβ1-3GalNAc), was found to react with the CgA from different animals by western blotting. Purified rabbit anti-bovine CgA antibody was also found to cross-react with the crude CgA preparations. On the basis of these findings, a sandwich ELISA was developed with ACA as the coating reagent and anti-bovine CgA antibody as the probing antibody. Using this method, concentration-dependent curves ranging from 0.003 μg/mL to 25 μg/mL and from 0.02 μg/mL to 25 μg/mL were obtained for bovine CgA and canine CgA, respectively. Similarly, concentration-dependent curves were obtained for the equine, swine, and dolphin crude CgA extracts. Thus, ACA is concluded to be a valuable reagent for CgA detection in crude extracts from different animal species, and for CgA isolation/purification.
authors: Laboratory of Veterinary Surgery, Division of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Japan. sayochan@vet.osakafu-u.ac.jp
authors: Hideo
authors: H
authors: Akiyoshi
authors: Shunji
authors: S
authors: Sugii
authors: Md A
authors: MA
authors: Nahid
authors: Katsuhito
authors: K
authors: Sone
authors: Toshiyuki
authors: T
authors: Tanaka
authors: Cao
authors: C
authors: Zheng
authors: Li
authors: L
authors: Yijyun
authors: Mica
authors: M
authors: Aoki
authors: Shigeo
authors: S
authors: Takenaka
authors: Terumasa
authors: T
authors: Shimada
authors: Junichiro
authors: J
authors: Shimizu
authors: Ken-Ichi
authors: K
authors: Kiyomiya
authors: Fumihito
authors: F
authors: Ohashi
mesh_terms: D000311:Adrenal Glands / Q000737:chemistry*; D027721:Amaranthus / Q000276:immunology*; D000818:Animals; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000276:immunology*; D015153:Blotting, Western; D002417:Cattle; D053379:Chromogranin A / Q000032:analysis* / Q000276:immunology; D004285:Dogs; D004289:Dolphins; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006736:Horses; D037121:Plant Lectins / Q000276:immunology*; D011817:Rabbits / Q000276:immunology; D013552:Swine; D014020:Tissue Extracts / Q000737:chemistry
chemical_list: D015295:Antigens, Tumor-Associated, Carbohydrate; D053379:Chromogranin A; D037121:Plant Lectins; D014020:Tissue Extracts; C016467:Thomsen-Friedenreich antigen