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Label-Free Nuclease Assay with Long-Term Stability
Rui Liu1, Jianyu Hu1, Yongxin Chen1
1Key Laboratory of Green Chemistry & Technology, Ministry of Education, College of Chemistry , Sichuan University , Chengdu 610064 , China.
This study introduces a stable nuclease assay using elemental mass spectrometry for detecting copper nanoparticles (CuNPs). This method overcomes signal instability, offering high sensitivity and long-term stability for enzyme detection.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Nanotechnology
Background:
- Enzyme detection is crucial in biological processes.
- DNA-templated copper nanoparticles (CuNPs) offer a label-free assay but suffer from poor signal stability.
- Developing stable and sensitive enzyme assays remains a challenge.
Purpose of the Study:
- To develop a long-term stable and highly sensitive nuclease assay.
- To overcome the signal instability issue of traditional DNA-templated CuNP assays.
- To utilize elemental mass spectrometry for enzyme detection.
Main Methods:
- Utilized elemental mass spectrometry for detecting copper nanoparticles (CuNPs).
- Employed DNA-templated CuNPs with elemental mass spectrometry detection.
- Investigated various experimental conditions including template length, concentrations, and reaction times.
Main Results:
- Achieved long-term signal stability for the nuclease assay, with signals unchanged for at least 18 days.
- Demonstrated high sensitivity using inductively coupled plasma mass spectrometry (ICPMS) 63Cu signal, with a detection limit of 0.029 U/mL for Exonuclease I (Exo I).
- Established a dynamic range of 0.1 U/mL to 20 U/μL for Exo I concentration.
Conclusions:
- The proposed elemental mass spectrometry-based nuclease assay offers superior signal stability compared to traditional fluorescence methods.
- The assay is highly sensitive and reliable, validated by spiked-recovery tests in cell medium.
- This method provides a robust platform for sensitive and stable enzyme detection.
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