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Development of a highly sensitive chemiluminescence enzyme immunoassay using enhanced luminol as substrate
Author(s) -
Tao Xiaoqi,
Wang Wenjun,
Wang Zhanhui,
Cao Xingyuan,
Zhu Jinghui,
Niu Lanlan,
Wu Xiaoping,
Jiang Haiyang,
Shen Jianzhong
Publication year - 2014
Publication title -
luminescence
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.428
H-Index - 45
eISSN - 1522-7243
pISSN - 1522-7235
DOI - 10.1002/bio.2544
Subject(s) - chemiluminescence , luminol , horseradish peroxidase , chemistry , detection limit , substrate (aquarium) , immunoassay , luminescent measurements , chromatography , enzyme , biochemistry , luminescence , materials science , oceanography , optoelectronics , antibody , immunology , biology , geology
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. Copyright © 2013 John Wiley & Sons, Ltd.

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