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Colorimetric detection of furin based on enhanced catalytic activity of G-quadruplex/hemin DNAzyme.
Shi, Liu; Wang, Lin; Yu, Xiaomeng; Kuang, Deqi; Huang, Yue; Yang, Nana; Yang, Jie; Li, Genxi.
Afiliación
  • Shi L; Center for Molecular Recognition and Biosensing, School of Life Sciences, Shanghai University, Shanghai, 200444, PR China.
  • Wang L; State Key Laboratory of Analytical Chemistry for Life Science, School of Life Sciences, Nanjing University, Nanjing, 210023, PR China.
  • Yu X; State Key Laboratory of Analytical Chemistry for Life Science, School of Life Sciences, Nanjing University, Nanjing, 210023, PR China.
  • Kuang D; State Key Laboratory of Analytical Chemistry for Life Science, School of Life Sciences, Nanjing University, Nanjing, 210023, PR China.
  • Huang Y; Department of Food Science and Engineering, College of Light Industry and Food Engineering, Nanjing Forestry University, Nanjing, 210037, PR China. Electronic address: huangyue@njfu.edu.cn.
  • Yang N; Department of Obstetrics and Gynecology, the First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, PR China. Electronic address: nanayang210@163.com.
  • Yang J; State Key Laboratory of Analytical Chemistry for Life Science, School of Life Sciences, Nanjing University, Nanjing, 210023, PR China.
  • Li G; Center for Molecular Recognition and Biosensing, School of Life Sciences, Shanghai University, Shanghai, 200444, PR China; State Key Laboratory of Analytical Chemistry for Life Science, School of Life Sciences, Nanjing University, Nanjing, 210023, PR China.
Anal Chim Acta ; 1323: 343070, 2024 Sep 22.
Article en En | MEDLINE | ID: mdl-39182972
ABSTRACT

BACKGROUND:

Rapid and sensitive colorimetric detection methods are crucial for diseases diagnosis, particularly those involving proteases like furin, which are implicated in various conditions, including cancer. Traditional detection methods for furin suffer from limitations in sensitivity and practicality for on-site detection, motivating the development of novel detection strategies. Therefore, developing a simple, enzyme-free, and rapid colorimetric analysis method with high sensitivity for furin detection is imperative.

RESULTS:

Herein, we have proposed a colorimetric method in this work for the first time to detect furin, leveraging the assembly of G-quadruplex/hemin DNAzyme with enhanced catalytic activity. Specifically, a peptide-DNA conjugate (PDC) comprising a furin-recognition peptide and flanking DNA sequences for signal amplification is designed to facilitate the DNAzyme assembly. Upon furin treatment, PDC cleavage triggers a cyclic catalytic hairpin assembly reaction to form the complementary double-stranded structures by hairpin 1 (HP1) and hairpin 2 (HP2), bringing the G-quadruplex sequence in HP1 closer to hemin on HP2. Moreover, the resulting G-quadruplex/hemin DNAzymes exhibit robust peroxidase-like activity, enabling the catalysis of the colorimetric reaction of ABTS2- for furin detection. Our method demonstrates high sensitivity, rapid response, and compatibility with complex sample matrices, achieving a detection limit as low as 1.1 pM.

SIGNIFICANCE:

The DNAzyme reported in this work exhibits robust catalytic activity, enabling high sensitivity and good efficiency for the detection. By eliminating the requirement for exogenous enzymes, our approach enables visual furin detection without expensive instrumentation and reagents, promising significant utility in biomedical and clinical diagnostic applications. Given the various design of peptide sequence and the programmability of DNA, it can be readily applied to analyzing other useful tumor biomarkers.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Colorimetría / ADN Catalítico / Furina / G-Cuádruplex / Hemina Límite: Humans Idioma: En Revista: Anal Chim Acta Año: 2024 Tipo del documento: Article Pais de publicación: Países Bajos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Colorimetría / ADN Catalítico / Furina / G-Cuádruplex / Hemina Límite: Humans Idioma: En Revista: Anal Chim Acta Año: 2024 Tipo del documento: Article Pais de publicación: Países Bajos