BASED ON QUARTZ CRYSTAL MICROBALANCE

Abstract

In this study, a genosensor based on quartz crystal microbalance QCM has been developed for the detection of single-stranded oligonucleotide specific to alpha-fetoprotein AFP gene. The quartz crystals were coated with polyethyleneimine PEI and then crosslinked with glutaraldehyde GA . The oligonucleotide probe was immobilized onto the PEI-GA film. The coatings were characterized by using atomic force microscopy. Hybridization reaction between probe and the complementary strand was monitored by the QCM system in real-time. This work demonstrated that QCM is a suitable tool for the real-time monitoring of hybridization on the crystal surface without using any label.

Keywords

• Piezoelectric Sensor, Alpha-fetoprotein, Quartz Crystal Microbalance, Resonance Frequency, Atomic Force Microscopy, Hybridization, Gene Sensor

ALPHA-FETOPROTEIN GENOSENSOR BASED ON QUARTZ CRYSTAL MICROBALANCE

Abstract

Bu çalışmada, alfa-fetoprotein AFP genine özgü tek zincirli oligonükleotit dizisinin belirlenmesine yönelik olarak geliştirilen, kuartz kristal mikrobalans KKM sistemine dayalı bir gen sensörü anlatılmıştır. Kuartz kristaller, poli etilenimin PEI polimeri ile kaplanmış, ardından glutaraldehit GA ile çapraz bağlama işlemi gerçekleştirilmiştir. Oligonükleotit probu, hazırlanan PEI/GA yüzeyi üzerine immobilize edilmiştir. Yüzeylerin kaplanması atomik kuvvet mikroskobu ile doğrulanmıştır. Tamamlayıcı ve prob dizileri arasında gerçekleşen hibridizasyon reaksiyonu, KKM sistemi ile eş-zamanlı olarak görüntülenmiştir. Bu çalışma ile, KKM sensörünün, kristal yüzeyi üzerinde gerçekleşen hibridizasyon reaksiyonunu, herhangi bir işaretleyici kullanmadan, eş-zamanlı olarak belirleyebildiği gösterilmiştir.

References

1. Ş. Şeker and Y.M. Elçin. Bioanalytical applications of piezoelectric sensors. In: Selimovic S, Ed. Nanopatterning and nanoscale devices for biological applications. USA: CRC Press, Taylor and Francis Group, (2014) 259-285.
2. T. Goda, Y. Maeda, and Y. Miyahara, Simultaneous monitoring of protein adsorption kinetics using a quartz crystal microbalance and field-effect transistor integrated device. Analytical Chemistry, 84/17 (2012) 7308-7314.
3. C. Crosson, and C. Rossi, Quartz crystal microbalance immunosensor for the quantification of immunoglobulin G in bovinemilk. Biosensors and Bioelectronics, 42 (2013) 453-459.
4. P. A. Rasheed, and N. Sandhyarani, Quartz crystal microbalance genosensor for sequence specific detection of attomolar DNA targets. Analytica Chimica Acta, 905 (2016) 134-139.
5. X. Guo, C.S. Lin, S.H. Chen, R. Ye, and V.C. Wu, A piezoelectric immunosensor for specific capture and enrichment of viable pathogens by quartz crystal microbalance sensor, followed by detection with antibody-functionalized Bioelectronics, 38/1 (2012) 177-183. gold
6. nanoparticles. Biosensors and
7. J. Telegdi, A. Shaban, and E. Kálmán, EQCM study of copper and iron corrosion inhibition in presence of organic inhibitors and biocides. Electrochimica Acta, 45/22 (2000) 3639-3647.
8. P. Skládal, Piezoelectric quartz crystal sensors applied for bioanalytical assays and characterization of affinity interactions. Journal of the Brazilian Chemical Society, 14/4 (2003) 491-502.

9. A. Kumar, Biosensors based on piezoelectric crystal detectors: theory and application. JOM-e, 52/10 (2000) 1-6.
10. G. Sauerbrey, Verwendung von Schwingquarzen zur Wägung dünner Schichten und zur Mikrowägung. Zeitschrift Physik, 155 (1959) 206– 212.
11. T.N. Ly, S. Park, and S.J. Park, Detection of HIV-1 antigen by quartz crystal microbalance using gold nanoparticles. Sensors and Actuators B: Chemical, 237 (2016) 452-458.
12. Y.K. Kim, S.I. Lim, Y.Y. Cho, S. Choi, J.Y. Song, and D.J. An, Detection of H3N2 canine influenza virus using a quartz crystal microbalance. Journal of Virological Methods, 208 (2014) 16-20.
13. D. Wang, G. Chen, H. Wang, W. Tang, W. Pan, N. Li, and F. Liu, A reusable quartz crystal microbalance biosensor for highly specific detection of single-base DNA mutation. Biosensors and Bioelectronics, 48 (2013) 276-280.
14. P.A. Rasheed, and N. Sandhyarani, Femtomolar level detection of BRCA1 gene using a gold nanoparticle labeled sandwich type DNA sensor. Colloids and Surfaces B: Biointerfaces, 117 (2014) 7-13.
15. J. Jin, W. Jiang, J. Yin, X. Ji, and P. Stagnaro, Plasma proteins adsorption mechanism on polyethylene-grafted poly (ethylene glycol) surface by quartz crystal microbalance with dissipation. Langmuir, 29/22 (2013) 6624-6633.
16. Z. Yan, M. Yang, Z. Wang, F. Zhang, J. Xia, G. Shi, L. Xia, Y. Li, Y. Xia, L. Xia, A label-free immunosensor for detecting common acute lymphoblastic leukemia antigen (CD10) based on gold nanoparticles by quartz crystal microbalance. Sensors and Actuators B: Chemical, 210 (2015) 248-253.
17. Y.J. Kim, M.M. Rahman, and J.J. Lee, Ultrasensitive and label-free detection of annexin A3 based on quartz crystal microbalance. Sensors and Actuators B: Chemical, 177 (2013) 172-177.
18. W. Tao, Y. Ai, S. Liu, C.W. Lun, and P.T. Yung, Determination of Alpha-Fetoprotein by a Microfluidic Miniature Quartz Crystal Microbalance. Analytical Letters, 48/6 (2015) 907-920.
19. K.N. Huang, C.Y. Shen, S.H. Wang, and C.H. Hung, Development of quartz crystal microbalance-based immunosensor for detecting alpha- fetoprotein. Instrumentation Science & Technology, 41/3 (2013) 311- 324.
20. X. Zhou, L. Liu, M. Hu, L. Wang, and J. Hu, Detection of hepatitis B virus by piezoelctric biosensor, Journal of Pharmaceutical and Biomedical Analysis 27 (2002) 341-345.
21. W.C. Tsai, and I.C. Lin, Development of a piezoelectric immunosensor for the detection of alpha-fetoprotein. Sensors and Actuators B: Chemical, 106/1 (2005) 455-460.
22. S. Tombelli, M. Minunni, A. Santucci, M.M. Spiriti, and M. Mascini, A DNA-based piezoelectric biosensor: Strategies for coupling nucleic acids to piezoelectric devices. Talanta, 68/3 (2006) 806-812.