A Label-Free, Mix-and-Detect ssDNA-Binding Assay Based on Cationic Conjugated Polymers

被引:8
作者
Zhang, Pengbo [1 ,2 ]
Zandieh, Mohamad [2 ]
Ding, Yuzhe [2 ]
Wu, Lyuyuan [2 ]
Wang, Xiaoyu [3 ]
Liu, Juewen [2 ]
Li, Zhengping [1 ]
机构
[1] Univ Sci & Technol Beijing, Sch Chem & Biol Engn, 30 Xueyuan Rd, Beijing 100083, Peoples R China
[2] Univ Waterloo, Waterloo Inst Nanotechnol, Dept Chem, 200 Univ Ave West, Waterloo, ON N2L 3G1, Canada
[3] Univ Sci & Technol Beijing, Sch Mat Sci & Engn, 30 Xueyuan Rd, Beijing 100083, Peoples R China
来源
BIOSENSORS-BASEL | 2023年 / 13卷 / 01期
基金
加拿大自然科学与工程研究理事会;
关键词
cationic conjugated polymer; fluorescence; label-free; ssDNA; biosensor; GOLD NANOPARTICLES; COLORIMETRIC DETECTION; DNA METHYLATION; FLUORESCENT; STRATEGIES; SEQUENCES;
D O I
10.3390/bios13010122
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The accurate, simple, and efficient measurement of the concentration of single-stranded DNA (ssDNA) is important for many analytical applications, such as DNA adsorption, biosensor design, and disease diagnosis, but it is still a challenge. Herein, we studied a cationic conjugated polymer (CCP)-based ssDNA assay taking advantage of the obvious fluorescence change of CCPs upon binding ssDNA. Poly(3-(3 '-N,N,N-triethylamino-1 '-propyloxy)-4-methyl-2,5-thiophene hydrochloride) (PMNT) achieved an apparent dissociation constant (K-d) of 57 +/- 4 nM for ssDNA, indicating a very high binding affinity between PMNT and ssDNA. This allowed us to develop a CCP-based ssDNA biosensor with a detection limit of 0.6 nM, similar to the fluorescence-dye-based method using SYBR Green I and SYBR Gold. Our CCP-based biosensor produced smaller differences among ssDNA samples with different base compositions. In addition, the existence of double-stranded DNA (dsDNA) at different concentrations did not interfere with the fluorescence of PMNT, indicating that our CCP-based biosensor was more suitable for the measurement of ssDNA. Compared with fluorescence-intensity-based quantification, our CCP system allowed ratiometric quantification, which made the calibration easier and more robust. We then applied our method to the quantification of ssDNA on AuNPs using both unmodified and thiolated ssDNA, and the accurate quantification of ssDNA was achieved without any fluorophore modification. This method provides an alternative approach for the measurement of ssDNA.
引用
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页数:11
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