Skip to main content
SpringerLink
Log in

Lysozyme aptasensor based on a glassy carbon electrode modified with a nanocomposite consisting of multi-walled carbon nanotubes, poly(diallyl dimethyl ammonium chloride) and carbon quantum dots

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

Abstract

An aptamer-based method is described for electrochemical determination of lysozyme. A glassy carbon electrode was modified with a nanocomposite composed of multi-walled carbon nanotubes, poly(diallyl dimethyl ammonium chloride), and carbon quantum dots. The composition of the nanocomposite (MWCNT/PDDA/CQD) warrants good electrical conductivity and a high surface-to-volume ratio. The lysozyme-binding aptamers were immobilized on the nanocomposite via covalent coupling between the amino groups of the aptamer and the carboxy groups of the nanocomposite. The modified electrode was characterized by electrochemical impedance spectroscopy, cyclic voltammetry and differential pulse voltammetry. The use of this nanocomposite results in a considerable enhancement of the electrochemical signal and contributes to improving sensitivity. Hexacyanoferrate was used as an electrochemical probe to study the dependence of the peak current on lysozyme concentration. In the presence of lysozyme, the interaction of lysozyme with immobilized aptamer results in a decrease of the peak current, best measured at +0.15 V vs. Ag/AgCl. A plot of peak current changes versus the logarithm of the lysozyme concentration is linear in the 50 fmol L−1 to 10 nmol L−1 concentration range, with a 12.9 fmol L−1 detection limit (at an S/N ratio of 3). The method is highly reproducible, specific and sensitive, and the electrode has a rapid response. It was applied to the determination of lysozyme in egg white, serum, and urine.

Schematic of a nanocomposite composed of multi-walled carbon nanotubes (MWCNTs), poly(diallyldimethyl ammonium chloride) (PDDA), and carbon quantum dots (CQDs) for use in a lysozyme aptasensor. The aptamer was immobilized on the surface, and bovine serum albumin (BSA) was applied to block the surface. The changes of peak current for the electrochemical probe hexacyanoferrate (Fe(CN)63−/4-) in the presence and absence of lysozyme was traced.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Setford SJ, White SF, Bolbot JA (2002) Measurement of protein using an electrochemical bi-enzyme sensor. Biosens Bioelectron 17:79–86. https://doi.org/10.1016/S0956-5663(01)00264-0

    Article  CAS  Google Scholar 

  2. Gianazza E, Tremoli E, Banfi C (2014) The selected reaction monitoring/multiple reaction monitoring-based mass spectrometry approach for the accurate quantitation of proteins: clinical applications in the cardiovascular diseases. Expert Rev Proteom 11:771–788. https://doi.org/10.1586/14789450.2014.947966

    Article  CAS  Google Scholar 

  3. Zhang LX, Cao YR, Xiao H, Liu XP, Liu SR, Meng QH, Fan L, Cao C (2016) Leverage principle of retardation signal in titration of double protein via chip moving reaction boundary electrophoresis. Biosens Bioelectron 77:284–291. https://doi.org/10.1016/j.bios.2015.09.001

    Article  CAS  Google Scholar 

  4. Yoo G, Bong JH, Kim S, Jose J, Pyun JC (2014) Microarray based on autodisplayed Ro proteins for medical diagnosis of systemic lupus erythematosus (SLE). Biosens Bioelectron 57:213–218. https://doi.org/10.1016/j.bios.2014.02.018

    Article  CAS  Google Scholar 

  5. Goda T, Higashi D, Matsumoto A, Hoshi T, Sawaguchi T, Miyahara Y (2015) Dual aptamer-immobilized surfaces for improved affinity through multiple target binding in potentiometric thrombin biosensing. Biosens Bioelectron 73:174–180. https://doi.org/10.1016/j.bios.2015.05.067

    Article  CAS  Google Scholar 

  6. Liu DY, Zhao Y, He XW, Yin XB (2011) Electrochemical aptasensor using the tripropylamine oxidation to probe intramolecular displacement between target and complementary nucleotide for protein array. Biosens Bioelectron 26:2905–2910. https://doi.org/10.1016/j.bios.2010.11.035

    Article  CAS  Google Scholar 

  7. Cheng AKH, Ge BX, HZ Y (2007) Aptamer-based biosensors for label-free voltammetric detection of lysozyme. Anal Chem 79:5158–5164. https://doi.org/10.1021/ac062214q

    Article  CAS  Google Scholar 

  8. Vasilescu A, Wang Q, Li M, Boukherroub R, Szunerits S (2016) Aptamer-based electrochemical sensing of lysozyme. Chem Aust 4:10–30. https://doi.org/10.3390/chemosensors4020010

    Google Scholar 

  9. Weth F, Schroeder T, Buxtorf UP, Lebensm Z (1988) Determination of lysozyme content in eggs and egg products using SDS-gel electrophoresis. Eur Food Res Technol 187:541–545. https://doi.org/10.1007/BF01042386

    CAS  Google Scholar 

  10. Daeschel MA, Musafija-Jeknic T, Wu Y, Bizzarri D, Villa A (2002) High-performance liquid chromatography analysis of lysozyme in wine. Am J Enol Vitic 53:154–157

    CAS  Google Scholar 

  11. Lacorn MGC, Haas-Lauterbach S, Immer U (2010) Sensitive lysozyme testing in red and white wine using the ridascreen fast lysozyme ELISA. Bulletin de l'OIV 83:507–511

    CAS  Google Scholar 

  12. Zhao Y, Chen H, Chen Q, Qi Y, Yang F, Tang J, He P, Zhang F (2014) Ultrasensitive and signal-on electrochemiluminescence aptasensor using the multi-tris(bipyridine) ruthenium(II)-β-cyclodextrin complexes. Chin J Chem 32(11):1161–1168. https://doi.org/10.1002/cjoc.201400511

    Article  CAS  Google Scholar 

  13. Vasilescu A, Gaspar S, Mihai I, Tache A, Litescu SC (2013) Development of a label-free aptasensor for monitoring the self-association of lysozyme. Analyst 138:3530–3537. https://doi.org/10.1039/C3AN00229B

    Article  CAS  Google Scholar 

  14. Ocana C, Hayat A, Mishra R, Vasilescu A, del Valle M, Marty J-L (2015) A novel electrochemical aptamer-antibody sandwich assay for lysozyme detection. Analyst (Cambridge, UK) 140(12):4148–4153. https://doi.org/10.1039/C5AN00243E

    Article  CAS  Google Scholar 

  15. Mir M, Vreeke M, Katakis I (2006) Different strategies to develop an electrochemical thrombin aptasensor. Electrochem Commun 8:505–511. https://doi.org/10.1016/j.elecom.2005.12.022

    Article  CAS  Google Scholar 

  16. Wang S, Hu X, Tan L, Liao Q, Chen Z (2016) Colorimetric detection of lysozyme based on its effect on the growth of gold nanoparticles induced by the reaction of chloroauric acid and hydroxylamine. Microchim Acta 183:3135–3141. https://doi.org/10.1007/s00604-016-1969-2

    Article  CAS  Google Scholar 

  17. Yang L, Fung C, Cho EJ, Ellington AD (2007) Real-time rolling circle amplification for protein detection. Anal Chem 79:3320–3329. https://doi.org/10.1021/ac062186b

    Article  CAS  Google Scholar 

  18. Zheng B, Cheng S, Liu W, Lam MHW, Liang H (2013) Small organic molecules detection based on aptamer-modified gold nanoparticles-enhanced quartz crystal microbalance with dissipation biosensor. Anal Biochem 438:144–149. https://doi.org/10.1016/j.ab.2013.03.030

    Article  CAS  Google Scholar 

  19. Villamizar RA, Maroto A, Rius FX, Inza I, Figueras MJ (2008) Fast detection of salmonella Infantis with carbon nanotube field effect transistors. Biosens Bioelectron 24:279–283. https://doi.org/10.1016/j.bios.2008.03.046

    Article  CAS  Google Scholar 

  20. Bahsi ZB, Buyukaksoy A, Olmezcan SM, Simsek F, Aslan MH, Oral AY (2009) A novel label-free optical biosensor using synthetic oligonucleotides from E. coli O157: H7: elementary sensitivity tests. Sensors 9:4890–4900. https://doi.org/10.3390/s90604890

    Article  CAS  Google Scholar 

  21. Gehring AG, SI T (2005) Enzyme-linked immunomagnetic electrochemical detection of live Escherichia coli O157: H7 in apple juice. Food Prot 68:146–149. https://doi.org/10.4315/0362-028X-68.1.146

    Article  CAS  Google Scholar 

  22. Heydari-Bafrooei E, Askari S (2017) Ultrasensitive aptasensing of lysozyme by exploiting the synergistic effect of gold nanoparticle-modified reduced graphene oxide and MWCNTs in a chitosan matrix. Microchim Acta 184:3405–3413. https://doi.org/10.1007/s00604-017-2356-3

    Article  CAS  Google Scholar 

  23. Iliuk AB, Hu L, Tao W (2011) Aptamer in bioanalytical applications. Anal Chem 83:4440–4452. https://doi.org/10.1021/ac201057w

    Article  CAS  Google Scholar 

  24. Fang S, Dong X, Ji H, Liu S, Yan F, Peng D, Zhang Z (2016) Electrochemical aptasensor for lysozyme based on a gold electrode modified with a nanocomposite consisting of reduced graphene oxide, cuprous oxide, and plasma-polymerized propargylamine. Microchim Acta 183:633–642. https://doi.org/10.1007/s00604-015-1675-5

    Article  CAS  Google Scholar 

  25. Ensafi AA, Jamei HR, Heydari-Bafrooei E, Rezaei B (2014) Development of a voltammetric procedure based on DNA interaction for sensitive monitoring of chrysoidine, a banned dye, in foods and textile effluents. Sensors Actuators B 202:224–231. https://doi.org/10.1016/j.snb.2014.05.001

    Article  CAS  Google Scholar 

  26. Xie D, Li C, Shangguan L, Qi H, Xue D, Gao Q, Zhang C (2014) Click chemistry-assisted self-assembly of DNA aptamer on gold nanoparticles-modified screen-printed carbon electrodes for label-free electrochemical aptasensor. Sensors Actuators B 192:558–564. https://doi.org/10.1016/j.snb.2013.11.038

    Article  CAS  Google Scholar 

  27. Shamsipur M, Farzin L, Tabrizi MA (2016) Ultrasensitive aptamer-based on-off assay for lysozyme using a glassy carbon electrode modified with gold nanoparticles and electrochemically reduced graphene oxide. Microchim Acta 183:2733–2743. https://doi.org/10.1007/s00604-016-1920-6

    Article  CAS  Google Scholar 

  28. Ensafi AA, Jamei HR, Heydari-Bafrooei E, Rezaei B (2016) Electrochemical study of quinone redox cycling: a novel application of DNA-based biosensors for monitoring biochemical reactions. Bioelectrochemistry 111:15–22. https://doi.org/10.1016/j.bioelechem.2016.04.008

    Article  CAS  Google Scholar 

  29. Xiao Y, Wang Y, Wu M, Ma X, Yang X (2013) Graphene-based lysozyme binding aptamer nanocomposite for label-free and sensitive lysozyme sensing. J Electroanal Chem 702:49–55. https://doi.org/10.1016/j.jelechem.2013.05.010

    Article  CAS  Google Scholar 

  30. Xu J, Wang Y, Hu S (2017) Nanocomposites of graphene and graphene oxides: synthesis, molecular functionalization and application in electrochemical sensors and biosensors. A review. Microchim Acta 184(1):1–44

    Article  CAS  Google Scholar 

  31. Gao X, Cui Y, Levenson M, Chung LW, Nie S (2004) In vivo cancer targeting and imaging with semiconductor quantum dots. Nat Biotechnol 22:969–976

    Article  CAS  Google Scholar 

  32. Lim SY, Shen W, Gao Z (2015) Carbon quantum dots and their applications. Chem Soc Rev 44:362–381. https://doi.org/10.1039/C4CS00269E

    Article  CAS  Google Scholar 

  33. Qu D, Zheng M, Du P, Zhou Y, Zhang L, Li D, Tan H, Zhao Z, Xie Z, Sun Z (2013) Highly luminescent S, N co-doped graphene quantum dots with broad visible absorption bands for visible light photocatalysts. Nano 5:12272–12277. https://doi.org/10.1039/C3NR04402E

    CAS  Google Scholar 

  34. Yang X, Yang X, Li Z, Li S, Han Y, Chen Y, Bu X, Su C, Xu H, Jiang Y, Lin Q (2015) Photoluminescent carbon dots synthesized by microwave treatment for selective image of cancer cells. J Colloid Interface Sci 456:1–6. https://doi.org/10.1016/j.jcis.2015.06.002

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors wish to thank the Center of Excellence in Sensor and Green Chemistry and Research Council of Isfahan University of Technology (IUT), and the Iranian Nanotechnology Initiative Council for their support.

Author information

Authors and Affiliations

  1. Department of Chemistry, Isfahan University of Technology, Isfahan, 84156-83111 I.R., Iran

    Behzad Rezaei, Hamid Reza Jamei & Ali Asghar Ensafi

Authors
  1. Behzad Rezaei
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hamid Reza Jamei
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ali Asghar Ensafi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Behzad Rezaei.

Ethics declarations

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

Electronic supplementary material

ESM 1

(DOCX 323 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Rezaei, B., Jamei, H.R. & Ensafi, A.A. Lysozyme aptasensor based on a glassy carbon electrode modified with a nanocomposite consisting of multi-walled carbon nanotubes, poly(diallyl dimethyl ammonium chloride) and carbon quantum dots. Microchim Acta 185, 180 (2018). https://doi.org/10.1007/s00604-017-2656-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s00604-017-2656-7

Keywords

Search

Navigation