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Microchimica Acta

, Volume 182, Issue 9–10, pp 1685–1691 | Cite as

Determination of prostate-specific antigen in serum samples using gold nanoparticle based amplification and lab-on-a-chip based amperometric detection

  • Yildiz Uludag
  • Güzin Köktürk
Original Paper

Abstract

We describe an electrode array and microfluidics based integrated biochip for the quantitation of the tumor marker prostate specific antigen (PSA). The surface of the chip was functionalized with a self-assembled monolayer of mercaptoundecanoic acid prior to the immobilization of the antibody against PSA. A flow of buffer or spiked human serum (75 %) and, subsequently, the detection antibody and gold nanoparticles (Au-NPs) modified with horseradish peroxidase were passed over the antibody-coated electrodes. This was followed by the injection of the substrate tetramethylbenzidine and simultaneous amperometry during the flow. This resulted in a real-time amperometric reading. The method has detection limits (LODs) of 0.2 ng∙L−1 in buffer and of 1 ng∙L−1 in 75 % human serum. The linear part of the calibration plot has an r2 of 0.97. These LODs are well below the clinical threshold level of 4 ng∙L−1. This assay is rapid (~15 min) which compares favorably with respect to conventional chronoamperometric analysis and to ELISA tests which require ~45 min. This new platform has a potential as an automatted point-of-care device for clinical use because it is likely to be applicable to numerous other clinical analytes for which appropriate antibodies are available.

Graphical Abstract

An electrode array and microfluidics based integrated biochip has been utilised for the real-time amperometric detection of the tumor marker prostate specific antigen (PSA).

Keywords

Amperometry Electrochemical sensing Biosensor Cancer biomarker PSA 

Notes

Acknowledgments

The project is supported by the Republic of Turkey Ministry of Development Infrastructure Grant (no: 2011 K120020) and BILGEM - TUBITAK (The Scientific and Technological Research Council of Turkey) (grant no: S569000). We gratefully acknowledge Dr. Zehra Ölçer from GYTE, Aylin Ersoy, Sinan Budak, Atike Demiralp, Hakkı Aktepe, Tugba Yurt, and Muammer Karadağ from BILGEM - TUBITAK for their contribution to the fabrication of the electrode arrays and sensor cassette.

Conflict of interests

The authors declare no competing financial interest.

References

  1. 1.
    Diamandis EP (1998) Prostate-specific antigen: its usefulness in clinical medicine. Trends Endocrinol Metab 9:310–316CrossRefGoogle Scholar
  2. 2.
    Brawer MK, Lange PH (1989) PSA in the screening, staging and follow-up of early-stage prostate cancer. A review of recent developments. World J Urol 7:7–11CrossRefGoogle Scholar
  3. 3.
    Healy DA, Hayes CJ, Leonard P, McKenna L, O’Kennedy R (2007) Biosensor developments: application to prostate-specific antigen detection. Trends Biotechnol 25:125–131CrossRefGoogle Scholar
  4. 4.
    Golberg A, Yarmush ML, Konry T (2013) Picoliter droplet microfluidic immunosorbent platform for point-of-care diagnostics of tetanus. Microchim Acta 180:855–860CrossRefGoogle Scholar
  5. 5.
    Konry T, Bale SS, Bhushan A, Shen K, Seker E, Polyak B, Yarmush M (2012) Particles and microfluidics merged: perspectives of highly sensitive diagnostic detection. Microchim Acta 176:251–269CrossRefGoogle Scholar
  6. 6.
    Sarkar P, Pal PS, Ghosh D, Setford SJ, Tothill IE (2002) Amperometric biosensors for detection of the prostate cancer marker (PSA). Int J Pharm 238:1–9CrossRefGoogle Scholar
  7. 7.
    Okuno J, Maehashi K, Kerman K, Takamura Y, Matsumoto K, Tamiya E (2007) Label-free immunosensor for prostate-specific antigen based on single-walled carbon nanotube array-modified microelectrodes. Biosens Bioelectron 22:2377–2381CrossRefGoogle Scholar
  8. 8.
    Li C, Curreli M, Lin H, Lei B, Ishikawa FN, Datar R, Cote RJ, Thompson ME, Zhou C (2005) Complementary detection of prostate-specific antigen using In 2O3 nanowires and carbon nanotubes. J Am Chem Soc 127:12484–12485CrossRefGoogle Scholar
  9. 9.
    Fernandez-Sanchez C, McNeil CJ, Rawson K, Nilsson O (2004) Disposable noncompetitive immunosensor for free and total prostate-specific antigen based on capacitance measurement. Anal Chem 76:5649–5656CrossRefGoogle Scholar
  10. 10.
    Choi JW, Kang DY, Jang YH, Kim HH, Min J, Oh BK (2008) Ultra-sensitive surface plasmon resonance based immunosensor for prostate-specific antigen using gold nanoparticle-antibody complex. Colloids Surf A 313–314:655–659CrossRefGoogle Scholar
  11. 11.
    Uludag Y, Tothill IE (2012) Cancer biomarker detection in serum samples using surface plasmon resonance and quartz crystal microbalance sensors with nanoparticle signal amplification. Anal Chem 84:5898–5904CrossRefGoogle Scholar
  12. 12.
    Ye Z, Tan M, Wang G, Yuan J (2004) Preparation, characterization, and time-resolved fluorometric application of silica-coated terbium(III) fluorescent nanoparticles. Anal Chem 76:513–518CrossRefGoogle Scholar
  13. 13.
    Hwang KS, Lee JH, Park J, Yoon DS, Park JH, Kim TS (2004) In-situ quantitative analysis of a prostate-specific antigen (PSA) using a nanomechanical PZT cantilever. Lab Chip 4:547–552CrossRefGoogle Scholar
  14. 14.
    Lee JH, Hwang KS, Park J, Yoon KH, Yoon DS, Kim TS (2005) Immunoassay of prostate-specific antigen (PSA) using resonant frequency shift of piezoelectric nanomechanical microcantilever. Biosens Bioelectron 20:2157–2162CrossRefGoogle Scholar
  15. 15.
    Wee KW, Kang GY, Park J, Kang JY, Yoon DS, Park JH, Kim TS (2005) Novel electrical detection of label-free disease marker proteins using piezoresistive self-sensing micro-cantilevers. Biosens Bioelectron 20:1932–1938CrossRefGoogle Scholar
  16. 16.
    Zhang B, Zhang X, Yan H, Xu S, Tang D, Fu W (2007) A novel multi-array immunoassay device for tumor markers based on insert-plug model of piezoelectric immunosensor. Biosens Bioelectron 23:19–25CrossRefGoogle Scholar
  17. 17.
    Uludag Y, Tothill IE (2010) Development of a sensitive detection method of cancer biomarkers in human serum (75 %) using a quartz crystal microbalance sensor and nanoparticles amplification system. Talanta 82:277–282CrossRefGoogle Scholar
  18. 18.
    Shah J, Wilkins E (2003) Electrochemical biosensors for detection of biological warfare agents. Electroanalysis 15:157–167CrossRefGoogle Scholar
  19. 19.
    Chikkaveeraiah BV, Mani V, Patel V, Gutkind JS, Rusling JF (2011) Microfluidic electrochemical immunoarray for ultrasensitive detection of two cancer biomarker proteins in serum. Biosens Bioelectron 26:4477–4483CrossRefGoogle Scholar
  20. 20.
    Zani A, Laschi S, Mascini M, Marrazza G (2011) A new electrochemical multiplexed assay for PSA cancer marker detection. Electroanalysis 23:91–99CrossRefGoogle Scholar
  21. 21.
    Lin YY, Wang J, Liu G, Wu H, Wai CM, Lin Y (2008) A nanoparticle label/immunochromatographic electrochemical biosensor for rapid and sensitive detection of prostate-specific antigen. Biosens Bioelectron 23:1659–1665CrossRefGoogle Scholar
  22. 22.
    Liu S, Zhang X, Wu Y, Tu Y, He L (2008) Prostate-specific antigen detection by using a reusable amperometric immunosensor based on reversible binding and leasing of HRP-anti-PSA from phenylboronic acid modified electrode. Clin Chim Acta 395:51–56CrossRefGoogle Scholar
  23. 23.
    Uludag Y, Olcer Z, Samil Sagiroglu M (2014) Design and characterisation of a thin-film electrode array with shared reference/counter electrodes for electrochemical detection. Biosens Bioelectron 57:85–90CrossRefGoogle Scholar
  24. 24.
    Olcer Z, Esen E, Muhammad T, Ersoy A, Budak S, Uludag Y (2014) Fast and sensitive detection of mycotoxins in wheat using novel electrode arrays and real-time electrochemical profiling. Biosens Bioelectron 62:163–169CrossRefGoogle Scholar
  25. 25.
    Uludag Y, Sagiroglu M, Ersoy A, Edis A, Budak S, Demiralp A (2014) Electrochemical sensor array and its casette, In PT 2014/3992, T.P. Office, ed. (Turkey)Google Scholar
  26. 26.
    Ding L, Bond AM, Zhai J, 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.
    Cui R, Huang H, Yin Z, Gao D, Zhu J (2008) Horseradish peroxidase-functionalized gold nanoparticle label for amplified immunoanalysis based on gold nanoparticles/carbon nanotubes hybrids modified biosensor. Biosens Bioelectron 23:1666–1673CrossRefGoogle Scholar
  28. 28.
    Wang Q, Tang H, Xie Q, Jia X, Zhang Y, Tan L, Yao S (2008) The preparation and characterization of poly(o-phenylenediamine)/gold nanoparticles interface for immunoassay by surface plasmon resonance and electrochemistry. Colloids Surf B 63:254–261CrossRefGoogle Scholar
  29. 29.
    Wang Z, Yang Y, Leng K, Li J, Zheng F, Shen G, Yu R (2008) A sequence-selective electrochemical DNA biosensor based on HRP-labeled probe for colorectal cancer DNA detection. Anal Lett 41:24–35CrossRefGoogle Scholar
  30. 30.
    Uludag Y, Hammond R, Cooper MA (2010) A signal amplification assay for HSV type 1 viral DNA detection using nanoparticles and direct acoustic profiling. J Nanobiotechnol 8:3CrossRefGoogle Scholar
  31. 31.
    Schwartz JJ, Quakea SR (2007) High density single molecule surface patterning with colloidal epitaxy. Appl Phys Lett 91Google Scholar
  32. 32.
    Panini NV, Messina GA, Salinas E, Fernandez H, Raba J (2008) Integrated microfluidic systems with an immunosensor modified with carbon nanotubes for detection of prostate specific antigen (PSA) in human serum samples. Biosens Bioelectron 23:1145–1151CrossRefGoogle Scholar
  33. 33.
    Zhang S, Du P, Li F (2007) Detection of prostate specific antigen with 3,4-diaminobenzoic acid (DBA)-H2O2-HRP voltammetric enzyme-linked immunoassay system. Talanta 72:1487–1493CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Wien 2015

Authors and Affiliations

  1. 1.The Scientific and Technological Research Council of Turkey (TUBITAK)-BILGEM-UEKAE-Bioelectronic Devices and Systems GroupGebzeTurkey

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