title: Development of a highly sensitive chemiluminescence enzyme immunoassay using enhanced luminol as substrate.
abstract: In this study, a high sensitivity chemiluminescence enzyme immunoassay (CLEIA) based on novel enhancers was developed. Under optimal conditions, we developed an enhanced chemiluminescence reaction (ECR) catalyzed by horseradish peroxidase (HRP-C) in the presence of 3-(10'-phenothiazinyl) propane-1-sulfonate (SPTZ) and 4-morpholinopyridine (MORP) as enhancers. The limit of detection of the newly prepared chemiluminescent cocktail for HRP was 0.33 pg/well, which is lower than that of commercial Super Signal substrate. The results showed that this novel chemiluminescent cocktail can significantly increase the light output of HRP-catalyzed ECR, which can be translated into a corresponding improvement in sensitivity. Similar improvements were observed in CLEIA for the determination of chloramphenicol in milk. In addition, the ECR of N-azoles as secondary enhancer was also presented.
authors.affiliation: Department of Pharmacology and Toxicology, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China.
authors.forename: Xiaoqi
authors.initials: X
authors.lastname: Tao
authors.forename: Wenjun
authors.initials: W
authors.lastname: Wang
authors.forename: Zhanhui
authors.initials: Z
authors.lastname: Wang
authors.forename: Xingyuan
authors.initials: X
authors.lastname: Cao
authors.forename: Jinghui
authors.initials: J
authors.lastname: Zhu
authors.forename: Lanlan
authors.initials: L
authors.lastname: Niu
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Wu
authors.forename: Haiyang
authors.initials: H
authors.lastname: Jiang
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Shen
mesh_terms: D000818:Animals; D002417:Cattle; D002701:Chloramphenicol / Q000032:analysis*; D005506:Food Contamination / Q000032:analysis; D006735:Horseradish Peroxidase / Q000737:chemistry*; D006861:Hydrogen Peroxide / Q000737:chemistry; D007124:Immunoenzyme Techniques / Q000295:instrumentation / Q000379:methods*; D049449:Luminescence; D008163:Luminescent Measurements / Q000295:instrumentation / Q000379:methods*; D008165:Luminol / Q000737:chemistry*; D008892:Milk / Q000737:chemistry*
chemical_list: D008165:Luminol; D002701:Chloramphenicol; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase
keywords: 3-(10'-phenothiazinyl) propane-1-sulfonate; 4-morpholinopyridine; chemiluminescence enzyme immunoassay; chloramphenicol; horseradish peroxidase