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.
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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.
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The authors declare no competing financial interest.
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Uludag, Y., Köktürk, G. Determination of prostate-specific antigen in serum samples using gold nanoparticle based amplification and lab-on-a-chip based amperometric detection. Microchim Acta 182, 1685–1691 (2015). https://doi.org/10.1007/s00604-015-1477-9
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DOI: https://doi.org/10.1007/s00604-015-1477-9