Abstract
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.
基金项目
Key-Area Research and Development Program of Guangdong Province(2022B1111020003)
Guangzhou Talents Program for Innovation and Entrepreneurship(2021-L010)
Foshan"Blue Ocean Talent Program"for Innovation and Entrepreneurship(2230032002063)
Lingnan Medical Research Center of Guangzhou University of Chinese Medicine and GIRM Biosafety(Guangzhou)Co.,Ltd.for the sup()
NETL
NSTL
万方数据