Developing a fluorescence substrate for HRP-based diagnostic assays with superiorities over the commercial ADHP

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外文摘要：The combination of horseradish peroxidase(HRP)and a fluorescence substrate has been attract-ing great interests in developing sensitive biochemical analysis and immunoassays.10-Acetyl-3,7-dihydroxyphenoxazine(ADHP or Amplex red)is the most sensitive fluorogenic substrate known for HRP in current market,however,it suffers from some drawbacks,such as non-specific reactivity to carboxylesterase and limited fluorescence stability.In the present study,a novel HRP substrate 10-cyclopropylcarbonyl-dichloro-dihydroxyphenoxazine(AR-2),has been prepared,which exhibited im-proved sensitivity than ADHP in sensing HRP.Moreover,the fluorescence of AR-2/HRP demonstrated im-proved tolerance to physiological relevant pH fluctuation as compared to ADHP/HRP.Successful detec-tion of uric acid/urate oxidase reaction indicated excellent application prospect of AR-2/HRP for monitor-ing H2O2-generating biochemical reactions.More interestingly,an enzyme-linked immunosorbent assay(ELISA)using AR-2 as the fluorescence reporter has been successfully used in detecting IgG against se-vere acute respiratory syndrome coronavirus 2(SARS-CoV-2)from human serum samples.Overall,AR-2 exhibits improved performances over the commercial ADHP,which will be an ideal alternative to ADHP in HRP-based fluorescence biochemical analysis and immunoassays.

外文关键词：

Fluorescence immunoassayHRP fluorescence substrateHydroresorufinAmplex red/ADHPSARS-CoV-2

作者：

Zhichao Zhou、Fuqian Chen、Xiaotong Xia、Dong Ye、Rong Zhou、Lei Li、Tao Deng、Zhenhua Ding、Fang Liu

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作者单位：

Department of Radiation Medicine,Guangdong Provincial Key Laboratory of Tropical Disease Research,School of Public Health,Southern Medical University,Guangzhou 510515,China

State Key Laboratory of Respiratory Disease,the First Affiliated Hospital of Guangzhou Medical University,Guangzhou Medical University,Guangzhou 510120,China

Artemisinin Research Center,Guangzhou University of Chinese Medicine,Guangzhou 510006,China

School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China

Guangzhou Laboratory,Bio-Island,Guangzhou 510030,China

Nanshan Pharmaceutical Innovation Research Institute of Guangdong Province,Foshan 528225,China

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基金：

Key-Area Research and Development Program of Guangdong ProvinceGuangzhou Talents Program for Innovation and EntrepreneurshipFoshan"Blue Ocean Talent Program"for Innovation and EntrepreneurshipLingnan Medical Research Center of Guangzhou University of Chinese Medicine and GIRM Biosafety(Guangzhou)Co.,Ltd.for the sup

项目编号：

2022B11110200032021-L0102230032002063

出版年：

2024

DOI：

10.1016/j.cclet.2023.108970

中国化学快报(英文版)

中国化学会

中国化学快报(英文版)

CSTPCD

影响因子：0.771

ISSN：1001-8417

年,卷(期)：2024.35(6)