Skip to main content
SpringerLink
Log in

A cationic conjugated polymer coupled with exonuclease I: application to the fluorometric determination of protein and cell imaging

  • Original Paper
  • Published:
Microchimica Acta Aims and scope Submit manuscript

Abstract

A strategy is described for the detection of protein by using a cationic fluorescent conjugated polymer coupled with exonuclease I (Exo I). Taking streptavidin (SA) as model protein, it is observed that Exo I can digest single-stranded DNA conjugated with biotin and carboxyfluorescein (P1) if SA is absent. This leads to the formation of small nucleotide fragments and to weak fluorescence resonance energy transfer (FRET) from the polymer to P1. If, however, SA is present, the high affinity of SA and biotin prevents the digestion of P1 by Exo I. This results in the sorption of P1 on the surface of the polymer through strong electrostatic interaction. Hence, efficient FRET occurs from the fluorescent polymer to the fluorescent label of P1. Fluorescence is measured at an excitation wavelength of 370 nm, and emission is measured at two wavelengths (530 and 425 nm). The ratio of the two intensities (I530/I425) is directly related to the concentration of SA. Under the optimal conditions, the assay has a detection limit of 1.3 ng·mL−1. The method was also applied to image the folate receptor in HeLa cells, thus demonstrating the versatility of this strategy.

A fluorometric strategy is described for protein detection and cell imaging based on a cationic conjugated polymer (PFP) coupled with exonuclease I (Exo I) trigged fluorescence resonance energy transfer (FRET).

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Scheme 1
Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Farka Z, Juřík T, Kovář D, Trnková L, Skládal P (2017) Nanoparticle-based immunochemical biosensors and assays: recent advances and challenges. Chem Rev 117:9973–10042

    Article  CAS  Google Scholar 

  2. Chen CH, Liu YF, Zheng ZH, Zhou GH, Ji XH, Wang HZ, He ZK (2015) A new colorimetric platform for ultrasensitive detection of protein and cancer cells based on the assembly of nucleic acids and proteins. Anal Chim Acta 880:1–7

    Article  CAS  Google Scholar 

  3. Zhuang YD, Chiang PY, Wang CW, Tan KT (2013) Environment-sensitive fluorescent turn-on probes targeting hydrophobic ligand-binding domains for selective protein detection. Angew Chem Int Ed 52:8124–8128

    Article  CAS  Google Scholar 

  4. Wang CK, Dong XY, Liu Q, Wang K (2015) Label-free colorimetric aptasensor for sensitive detection of ochratoxin a utilizing hybridization chain reaction. Anal Chim Acta 860:83–88

    Article  CAS  Google Scholar 

  5. Meng HM, Liu H, Kuai H, Peng R, Mo L, Zhang XB (2016) Aptamer-integrated DNA nanostructures for biosensing, bioimaging and cancer therapy. Chem Soc Rev 45:2583–2602

    Article  CAS  Google Scholar 

  6. Su S, Sun HF, Cao WF, Chao J, Peng HZ, Zuo XL, Yuwen LH, Fan CH, LH W (2016) Dual-target electrochemical biosensing based on DNA structural switching on gold nanoparticle-decorated MoS2 Nanosheets. ACS Appl Mater Interfaces 8:6826–6833

    Article  CAS  Google Scholar 

  7. Ye X, Shi H, He XX, Wang KM, He DG, Yan LA, Xu FZ, Lei YL, Tang JL, Yu YR (2015) Iodide-responsive cu−au nanoparticle-based colorimetric platform for ultrasensitive detection of target cancer cells. Anal Chem 87:7141–7147

    Article  CAS  Google Scholar 

  8. Freeman R, Girsh J, Fang-ju Jou A, Annie Ho JA, Hug T, Dernedde J, Willner I (2012) Optical aptasensors for the analysis of the vascular endothelial growth factor (VEGF). Anal Chem 84:6192–6198

    Article  CAS  Google Scholar 

  9. Wu Z, Zhen Z, Jiang JH, Shen GL, Yu RQ (2009) Terminal protection of small-molecule-linked DNA for sensitive electrochemical detection of protein binding via selective carbon nanotube assembly. J Am Chem Soc 131:12325–12332

    Article  CAS  Google Scholar 

  10. Zhou GH, Zhang X, Ji XH, He ZK (2013) Ultrasensitive detection of small molecule-protein interaction via terminal protection of small molecule linked DNA and Exo III-aided DNA recycling amplification. Chem Commun 49:8854–8856

    Article  CAS  Google Scholar 

  11. Ni JC, Wang QX, Yang WQ, Zhao MM, Zhang Y, Guo LH, Q B, Li ZY, Yang HH (2016) Immobilization free electrochemical biosensor for folate receptor in cancer cells based on terminal protection. Biosens Bioelectron 86:496–501

  12. He Y, Jiao B (2016) Detection of biotin-streptavidin interactions via terminal protection of small molecule linked DNA and the formation of fluorescent DNA-templated silver nanoclusters. Microchim Acta 183:3183–3189

    Article  CAS  Google Scholar 

  13. Li P, Wang L, Zhu J, YS W, Jiang W (2015) Label-free and dual-amplified detection of protein via small molecule-ligand linked DNA and a cooperative DNA machine. Biosens Bioelectron 72:107–113

    Article  CAS  Google Scholar 

  14. Li JS, Deng T, Chu X, Yang RH, Jiang JH, Shen GL, Yu R (2010) Rolling circle amplification combined with gold nanoparticle aggregates for highly sensitive identification of single-nucleotide polymorphisms. Anal Chem 82:2811–2816

    Article  CAS  Google Scholar 

  15. Zhao J, SS H, Cao Y, Zhang B, Li GX (2015) Electrochemical detection of protein based on hybridization chain reaction-assisted formation of copper nanoparticles. Biosens Bioelectron 66:327–331

    Article  CAS  Google Scholar 

  16. Cao Y, Zhu S, JC Y, Zhu XJ, Yin YM, Li GX (2012) Protein detection based on small molecule-linked DNA. Anal Chem 84:4314–4320

    Article  CAS  Google Scholar 

  17. Cao JP, Wang W, Bo B, Mao XX, Wang KM, Zhu XL (2017) A dual-signal strategy for the solid detection of both small molecules and proteins based on magnetic separation and highly fluorescent copper nanoclusters. Biosens Bioelectron 90:534–541

    Article  CAS  Google Scholar 

  18. Wang LJ, Zhang Y, Zhang CY (2014) A target-triggered exponential amplification-based DNAzyme biosensor for ultrasensitive detection of folate receptors. Chem Commun 50:15393–15396

    Article  CAS  Google Scholar 

  19. LJ O, Wang HB, Chu X (2013) Terminal protection of small-molecule-linked DNA for sensitive fluorescence detection of protein binding based on nucleic acid amplification. Analyst 138:7218–7223

    Article  Google Scholar 

  20. Duan XR, Liu LB, Feng FD, Wang S (2010) Cationic conjugated polymers for optical detection of DNA methylation, lesions, and single nucleotide polymorphisms. Acc Chem Res 43:260–270

    Article  CAS  Google Scholar 

  21. Liang J, Li K, Liu B (2013) Visual sensing with conjugated polyelectrolytes. Chem Sci 4:1377–1394

    Article  CAS  Google Scholar 

  22. Feng LH, Zhu CL, Yuan HX, Liu LB, Lv FT, Wang S (2013) Conjugated polymer nanoparticles: preparation, properties, functionalization and biological applications. Chem Soc Rev 42:6620–6633

    Article  CAS  Google Scholar 

  23. Ho HA, Najari A, Leclerc M (2008) Optical detection of DNA and proteins with cationic polythiophenes. Acc Chem Res 41:168–178

    Article  CAS  Google Scholar 

  24. Xing XJ, Liu XG, He Y, Lin Y, Zhang CL, Tang HW, Pang DW (2013) Amplified fluorescent sensing of DNA using graphene oxide and a conjugated cationic polymer. Biomacromolecules 14:117–123

    Article  CAS  Google Scholar 

  25. Liu B, Bazan GC (2004) Homogeneous fluorescence-based DNA detection with water-soluble conjugated polymers. Chem Mater 16:4467–4476

    Article  CAS  Google Scholar 

  26. Chen C, Min W, Liu Y, Xu E, Wei W, Zhang Y, Liu S (2017) Visual and fluorometric determination of telomerase activity by using a cationic conjugated polymer and fluorescence resonance energy transfer. Microchim Acta 184:3453–3460

    Article  CAS  Google Scholar 

  27. Wu Y, Tan Y, JT W, Chen SY, Chen YZ, Zhou XW, Jang YY, Tan CY (2015) Fluorescence array-based sensing of metal ions using conjugated polyelectrolytes. ACS Appl Mater Interfaces 7:6882–6888

    Article  CAS  Google Scholar 

  28. Wang C, Tang YL, Liu Y, Guo Y (2014) Water-soluble conjugated polymer as a platform for adenosine deaminase sensing based on fluorescence resonance energy transfer technique. Anal Chem 86:6433–6438

    Article  CAS  Google Scholar 

  29. Feng FD, Tang YL, He F, MH Y, Duan XR, Wang S, Li YL, Zhu DB (2007) Cationic conjugated polymer/DNA complexes for amplified fluorescence assays of nucleases and methyltransferases. Adv Mater 19:3490–3495

    Article  CAS  Google Scholar 

  30. Stork M, Gaylord BS, Heeger AJ, Bazan GC (2002) Energy transfer in mixtures of water- soluble oligomers: effect of charge, aggregation, and surfactant complexation. Adv Mater 14:361–366

    Article  CAS  Google Scholar 

  31. Xiang X, Shi JB, Huang FH, Zheng MM, Deng QC, JQ X (2015) MoS2 nanosheet-based fluorescent biosensor for protein detection via terminal protection of small-molecule-linked DNA and exonuclease III-aided DNA recycling amplification. Biosens Bioelectron 74:227–232

    Article  CAS  Google Scholar 

  32. Jiang H, Xu G, Sun YM, Zheng WW, Zhu XX, Wang BJ, Zhang XJ, Wang GF (2015) A "turn-on" silver nanocluster based fluorescent sensor for folate receptor detection and cancer cell imaging under visual analysis. Chem Commun 5:11810–11813

    Article  Google Scholar 

  33. Liu QL, SH X, Niu CX, Li MF, He DC, ZL L, Liu M, Na N, Huang F, Jiang H, Ouyang J (2015) Distinguish cancer cells based on targeting turn-on fluorescence imaging by folate functionalized green emitting carbon dots. Biosens Bioelectron 64:119–125

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the National Natural Science Foundation of China (21505114), (31601438) and Doctorial Starting Fund of Xinxiang Medical University.

Author information

Authors and Affiliations

  1. School of Pharmacy, Xinxiang Medical University, Xinxiang, Henan, 453003, People’s Republic of China

    Yufei Liu, Huijuan Yan & Jingfang Shangguan

  2. Department of toxicology, School of Public Health, Xinxiang Medical University, Xinxiang, Henan, 453003, People’s Republic of China

    Liyun Gao

  3. Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan, Hubei, 430000, People’s Republic of China

    Zhen Zhang & Xia Xiang

Authors
  1. Yufei Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Liyun Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Huijuan Yan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jingfang Shangguan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Zhen Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xia Xiang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Yufei Liu or Xia Xiang.

Ethics declarations

The author(s) declare that they have no competing interests.

Electronic supplementary material

ESM 1

(DOC 1136 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Liu, Y., Gao, L., Yan, H. et al. A cationic conjugated polymer coupled with exonuclease I: application to the fluorometric determination of protein and cell imaging. Microchim Acta 185, 118 (2018). https://doi.org/10.1007/s00604-017-2661-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s00604-017-2661-x

Keywords

Search

Navigation