Label-free detection of histone based on cationic conjugated polymer-mediated fluorescence resonance energy transfer.

Journal: Talanta
PMID: 29332793

Abstract

A simple and homogeneous histone assay is developed based on histone-induced DNA compressing coupled with cationic conjugated polymer (CCP)-mediated fluorescence resonance energy transfer (FRET). In this strategy, the CCP serves as the FRET donor and SYBR Green I (SG), which can strongly fluoresce not at its free state but after intercalated into the double stranded calf thymus DNA (dsDNA), serves as the acceptor of FRET. In the absence of histone, the dsDNA-SG and CCP combine with each other through electrostatic interaction and the strong FRET from CCP to SG occurs due to the overlapping between the fluorescent emitting spectrum of the CCP and the absorption spectrum of SG. Upon the introduction of histone, the formed compact complex of histone/dsDNA will lead to the compression of dsDNA structure and prevent SG binding to dsDNA and fluorescing, which gives rise to a significant decrease of FRET efficiency between CCP and SG. Thus, the quantitative analysis of histone is realized by monitoring the change of FRET ratio, namely, the intensity ratio of the two emission bands of CCP and SG. Due to the light harvesting and fluorescence amplification properties of CCP, high sensitivity is achieved with a low detection limit of 0.74ng/mL histone. This strategy provides a simple, homogeneous and sensitive strategy for histone analysis in the study of histone-related biological processes.

Authors

  • Xiaozhen Lu
    Key Laboratory of Medicinal Chemistry and Molecular Diagnosis, Ministry of Education, College of Chemistry and Environmental Science, Hebei University, Baoding 071002, Hebei Province, PR China.
  • Hongxia Jia
    Key Laboratory of Medicinal Chemistry and Molecular Diagnosis, Ministry of Education, College of Chemistry and Environmental Science, Hebei University, Baoding 071002, Hebei Province, PR China.
  • Xiaohua Yan
    Key Laboratory of Medicinal Chemistry and Molecular Diagnosis, Ministry of Education, College of Chemistry and Environmental Science, Hebei University, Baoding 071002, Hebei Province, PR China.
  • Jingsha Wang
    Key Laboratory of Medicinal Chemistry and Molecular Diagnosis, Ministry of Education, College of Chemistry and Environmental Science, Hebei University, Baoding 071002, Hebei Province, PR China.
  • Yucong Wang
    Key Laboratory of Medicinal Chemistry and Molecular Diagnosis, Ministry of Education, College of Chemistry and Environmental Science, Hebei University, Baoding 071002, Hebei Province, PR China. Electronic address: wangyucong@hbu.edu.cn.
  • Chenghui Liu
    Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an 710062, Shaanxi Province, PR China. Electronic address: liuch@snnu.edu.cn.

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