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Microchimica Acta

, Volume 182, Issue 11–12, pp 1917–1924 | Cite as

Electrochemical sandwich immunoassay for the peptide hormone prolactin using an electrode modified with graphene, single walled carbon nanotubes and antibody-coated gold nanoparticles

  • Shengqiang Li
  • Yurong Yan
  • Liang Zhong
  • Ping Liu
  • Ye Sang
  • Wei Cheng
  • Shijia DingEmail author
Original Paper

Abstract

We describe a new kind of electrochemical immunoassay for the peptide hormone prolactin. A glassy carbon electrode (GCE) was modified with a hybrid material consisting of graphene, single walled carbon nanotubes and gold nanoparticles (AuNPs) in a chitosan (CS) matrix. The graphene and the single wall carbon nanotubes were first placed on the GCE, and the AuNPs were then electrodeposited on the surface by cyclic voltammetry. This structure results in a comparably large surface for immobilization of the capturing antibody (Ab1). The modified electrode was used in a standard sandwich-type of immunoassay. The secondary antibody (Ab2) consisted of AuNPs with immobilized Ab2 and modified with biotinylated DNA as signal tags. Finally, alkaline phosphatase was bound to the biotinylated DNA-AuNPs-Ab2 conjugate via streptavidin chemistry. The enzyme catalyzes the hydrolysis of the α-naphthyl phosphate to form α-naphthol which is highly electroactive at an operating voltage as low as 180 mV (vs. Ag/AgCl). The resulting immunoassay exhibits high sensitivity, wide linear range (50 to 3200 pg∙mL‾1), low detection limit (47 pg∙mL‾1), acceptable selectivity and reproducibility. The assay provides a pragmatic platform for signal amplification and has a great potential for the sensitive determination of antigens other than prolactine.

Graphical Abstract

The immunoassay for prolactin is based on a glassy carbon electrode modified with SWCNTs, graphene and antibody-coated gold nanoparticles, and a secondary antibody conjugated to other gold nanoparticles via a biotinylated DNA linker

Keywords

Electrochemical biosensor Immunosensor Graphene Single wall carbon nanotubes Gold nanoparticles Prolactin Chitosan Alkaline phosphatase 

Notes

Acknowledgments

This work was funded by the National Natural Science Foundation of China (81371904) and (81101638), the Natural Science Foundation Project of CQ (CSTC2013jjB10019).

Supplementary material

604_2015_1528_MOESM1_ESM.doc (6.7 mb)
ESM 1 (DOC 6900 kb)

References

  1. 1.
    Freeman ME, Kanyicska B, Lerant A, Nagy G (2000) Prolactin: structure, function, and regulation of secretion. Physiol Rev 80(4):1523–1631Google Scholar
  2. 2.
    Moreno-Guzmán M, González-Cortés A, Yáñez-Sedeño P, Pingarrón JM (2011) A disposable electrochemical immunosensor for prolactin involving affinity reaction on streptavidin-functionalized magnetic particles. Anal Chim Acta 692(1):125–130CrossRefGoogle Scholar
  3. 3.
    Sinha YN (1995) Structural variants of prolactin: occurrence and physiological significance. Endocr Rev 16(3):354–369CrossRefGoogle Scholar
  4. 4.
    Fahie-Wilson M, Smith TP (2013) Determination of prolactin: the macroprolactin problem. Best Pract Res Clin Endocrinol Metab 27(5):725–742CrossRefGoogle Scholar
  5. 5.
    Banerjee S, Paul P, Talib VJ (2004) Serum prolactin in seizure disorders. Indian Pediatr 41(8):827–831Google Scholar
  6. 6.
    Bole-Feysot C, Goffin V, Edery M, Binart N, Kelly PA (1998) Prolactin (PRL) and its receptor: actions, signal transduction pathways and phenotypes observed in PRL receptor knockout mice. Endocr Rev 19(3):225–268CrossRefGoogle Scholar
  7. 7.
    Smith TP, Suliman AM, Fahie-Wilson MN, McKenna TJ (2002) Gross variability in the detection of prolactin in sera containing Big prolactin (macroprolactin) by commercial immunoassays. J Clin Endocrinol Metab 87(12):5410–5415CrossRefGoogle Scholar
  8. 8.
    Roy KS, Prakash BS (2007) Development and validation of a simple, sensitive enzyme immunoassay (EIA) for quantification of prolactin in buffalo plasma. Theriogenology 67(3):572–579CrossRefGoogle Scholar
  9. 9.
    Mondal M, Rajikhowa C, Prakash BS (2007) Development and validation of a highly sensitive economic enzymeimmunoassay for prolactin determination in blood plasma of mithun (Bos frontalis) and its application during milk let down and cyclicity. Anim Reprod Sci 99(1):182–195CrossRefGoogle Scholar
  10. 10.
    Kudryavtsev AN, Krasitskaya VV, Petunin AI, Burakov AY, Frank LA (2012) Simultaneous bioluminescent immunoassay of serum total and IgG-bound prolactins. Anal Chem 84(7):3119–3124CrossRefGoogle Scholar
  11. 11.
    Rojanasakul A, Udomsubpayakul U, Chinsomboon S (1994) Chemiluminescence immunoassay versus radioimmunoassay for the measurement of reproductive hormones. Int J Gynaecol Obstet 45(2):141–146CrossRefGoogle Scholar
  12. 12.
    Chikkaveeraiah BV, Bhirde AA, Morgan NY, Eden HS, Chen XY (2012) Electrochemical immunosensors for detection of cancer protein biomarkers. ACS Nano 6(8):6546–61CrossRefGoogle Scholar
  13. 13.
    Nie H, Liu S, Yu R, Jiang J (2009) Phospholipid-coated carbon nanotubes as sensitive electrochemical labels with controlled-assembly-mediated signal transduction for magnetic separation immunoassay. Angew Chem Int Ed 48(52):9862–6CrossRefGoogle Scholar
  14. 14.
    Wang J, Lin YH (2008) Functionalized carbon nanotubes and nanofibers forbiosensing applications. TrAC Trends Anal Chem 27(7):619–626CrossRefGoogle Scholar
  15. 15.
    Chuvilin A, Bichoutskaia E, Gimenez-Lopez MC, Chamberlain TW, Rance GA, Kuganathan N, Biskupek J, Kaiser U, Khlobystov AN (2011) Self-assembly of a sulphur-terminated graphene nanoribbon within a single-walled carbon nanotube. Nat Mater 10(9):687–692CrossRefGoogle Scholar
  16. 16.
    Chen X, Zhu J, Xi Q, Yang WS (2012) A high performance electrochemical sensor for acetaminophen based on single-walled carbon nanotube–graphene nanosheet hybrid films. Sensors Actuators B Chem 161(1):648–654CrossRefGoogle Scholar
  17. 17.
    Gan T, Hu SS (2011) Electrochemical sensors based on graphene materials. Microchim Acta 175(1–2):1–19CrossRefGoogle Scholar
  18. 18.
    Lu JJ, Liu SQ, Ge SG, Yan M, Yu JH, Hu XT (2012) Ultrasensitive electrochemical immunosensor based on Au nanoparticles dotted carbon nanotube–graphene composite and functionalized mesoporous materials. Biosens Bioelectron 33(1):29–35CrossRefGoogle Scholar
  19. 19.
    Zhu J, Chauhan DS, Shan D, Wu XY, Zhang GY, Zhang XJ (2014) Ultrasensitive determination of hydrazine using a glassy carbon electrode modified with Pyrocatechol Violet electrodeposited on single walled carbon nanotubes. Microchim Acta 181(7–8):813–820CrossRefGoogle Scholar
  20. 20.
    Zhang L, Li C, Zhao D, Wu T, Nie G (2014) An electrochemical immunosensor for the tumor marker α-fetoprotein using a glassy carbon electrode modified with a poly (5-formylindole), single-wall carbon nanotubes, and coated with gold nanoparticles and antibody. Microchim Acta 81(13–14):1601–1608CrossRefGoogle Scholar
  21. 21.
    Khorsand F, Darziani Azizi M, Naeemy A, Larijani B, Omidfar K (2013) An electrochemical biosensor for 3-hydroxybutyrate detection based on screen-printed electrode modified by coenzyme functionalized carbon nanotubes. Mol Biol Rep 40(3):2327–2334CrossRefGoogle Scholar
  22. 22.
    Liu Y, Liu Y, Feng HB, Wu YM, Joshi L, Zeng XQ, Li JH (2012) Layer-by-layer assembly of chemical reduced graphene and carbon nanotubes for sensitive electrochemical immunoassay. Biosens Bioelectron 35:63–68CrossRefGoogle Scholar
  23. 23.
    Zhao MQ, Liu XF, Zhang Q, Tian GL, Huang JQ, Zhu WC, Wei F (2012) Graphene/single-walled carbon nanotube hybrids: one-step catalytic growth and applications for high-rate Li–S batteries. ACS Nano 6(12):10759–10769Google Scholar
  24. 24.
    Cheng Q, Tang J, Ma J, Zhang H, Norio SY, Qin LC (2011) Graphene and carbon nanotube composite electrodes for supercapacitors with ultra-high energy density. Phys Chem Chem Phys 13(39):17615–17624CrossRefGoogle Scholar
  25. 25.
    Huang KJ, Li J, Liu YM, Cao X, Yu S, Yu M (2012) Disposable immunoassay for hepatitis B surface antigen based on a graphene paste electrode functionalized with gold nanoparticles and a Nafion-cysteine conjugate. Microchim Acta 177(3–4):419–426CrossRefGoogle Scholar
  26. 26.
    Ding L, Bond AM, Zhai JP, Zhang J (2013) Utilization of nanoparticle labels for signal amplification in ultrasensitive electrochemical affinity biosensors: a review. Anal Chim Acta 797:1–12CrossRefGoogle Scholar
  27. 27.
    Han E, Ding L, Ju HX (2011) Highly sensitive fluorescent analysis of dynamic glycan expression on living cells using glyconanoparticles and functionalized quantum dots. Anal Chem 83(18):7006–7012CrossRefGoogle Scholar
  28. 28.
    Shao K, Wang J, Jiang XC, Shao F, Li TT, Ye SY, Chen L, Han HY (2014) Stretch − stowage − growth strategy to fabricate tunable TriplyAmplified electrochemiluminescence immunosensor for ultrasensitive detection of pseudorabies virus antibody. Anal Chem 86:5749–5757CrossRefGoogle Scholar
  29. 29.
    Song C, Xie GM, Wang L, Liu LZ, Tian G, Xiang H (2014) DNA-based hybridization chain reaction for an ultrasensitive cancer marker EBNA-1 electrochemical immunosensor. Biosens Bioelectron 58:68–74CrossRefGoogle Scholar
  30. 30.
    Zhang B, Liu BQ, Tang DP, Niessner R, Chen GN, Knopp D (2012) DNA-based hybridization chain reaction for amplified bioelectronic signal and ultrasensitive detection of proteins. Anal Chem 84:5392–5399CrossRefGoogle Scholar
  31. 31.
    Tang J, Tang DP, Su BL, Huang JX, Qiu B, Chen GN (2011) Enzyme-free electrochemical immunoassay with catalytic reduction of p-nitrophenol and recycling of p-aminophenol using gold nanoparticles-coated carbon nanotubes as nanocatalysts. Biosens Bioelectron 26:3219–3226CrossRefGoogle Scholar
  32. 32.
    Kim KS, Zhao Y, Jang H, Lee SY, Kim JM, Kim KS, Ahn JH, Kim P, Choi JY, Hong BH (2009) Large-scale pattern growth of graphene films for stretchable transparent electrodes. Nature 457(7230):706–710CrossRefGoogle Scholar
  33. 33.
    Guo SJ, Wang EK (2007) Synthesis and electrochemical applications of gold nanoparticles. Anal Chim Acta 598(2):181–192CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Wien 2015

Authors and Affiliations

  • Shengqiang Li
    • 1
  • Yurong Yan
    • 1
  • Liang Zhong
    • 1
  • Ping Liu
    • 3
  • Ye Sang
    • 1
  • Wei Cheng
    • 1
    • 2
  • Shijia Ding
    • 1
    Email author
  1. 1.Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), Department of Laboratory MedicineChongqing Medical UniversityChongqingChina
  2. 2.Molecular Oncology and Epigenetics Laboratorythe First Affiliated Hospital of Chongqing Medical UniversityChongqingChina
  3. 3.Bioscience (Tianjin) Diagnostic Technology Co.LtdTianjinChina

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