Development of a highly sensitive chemiluminescence enzyme immunoassay using enhanced luminol as substrate |
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| Authors: | Xiaoqi Tao Wenjun Wang Zhanhui Wang Xingyuan Cao Jinghui Zhu Lanlan Niu Xiaoping Wu Haiyang Jiang Jianzhong Shen |
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| Institution: | 1. Department of Pharmacology and Toxicology, College of Veterinary Medicine, China Agricultural University, Beijing, China;2. Beijing WDWK Biotech Co., Ltd., Beijing, China |
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| 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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| Keywords: | 3‐(10'‐phenothiazinyl) propane‐1‐sulfonate 4‐morpholinopyridine horseradish peroxidase chemiluminescence enzyme immunoassay chloramphenicol |
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