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

作者：Akiyoshi Hideo^*; Sugii Shunji; Nahid Md A; Sone Katsuhito; Tanaka Toshiyuki; Zheng Cao; Yijyun Li; Aoki Mica; Takenaka Shigeo; Shimada Terumasa; Shimizu Junichiro; Kiyomiya Ken ichi; Ohashi Fumihito

来源：Veterinary Immunology and Immunopathology, 2011, 144(3-4): 255-258.

DOI：10.1016/j.vetimm.2011.08.024

• Chromogranin A
• Amaranthus caudatus
• Lectin

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 (Gal beta 1-3GalNAc), 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 mu g/mL to 25 mu g/mL and from 0.02 mu g/mL to 25 mu g/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.

• 出版日期2011-12-15