Abstract
A method for using a hyperbranched polymer (HBP) as a bridge to link multiple secondary antibodies at HBP branches to amplify the detection response signal on a quartz crystal microbalance (QCM)-based sandwich-type immunosensor is reported. Carboxyl groups were prepared at multiple branches of HBP to make possible chemical binding between HBP and secondary antibodies via the carboxyl-amine reaction. The total mass of HBP and its linked multiple secondary antibodies were used to enhance the signal on a QCM chip in comparison with a simple sandwich-type immune reaction. By contrast, the proposed method could cause one antigen to analogously react with multiple secondary antibodies as a result of the branch structure of HBP. The strategy of using HBP as a bridge to link multiple secondary antibodies succeeded in quantitatively detecting the hepatitis B surface antigen (HBsAg). By employing demonstrated HBP bridge-linking, the frequency shift on a QCM chip was approximately 5 times greater than conventional methods without modification at secondary antibodies. The limit of detection of HBsAg was achieved as 2.0 ng mL−1, lower than most of the values recorded in the literature measured by the QCM technique. Taking into account the general chemical interaction of immunoreaction, this method has the potential to amplify the signal in sensing many other analytes of interest.
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Bayramoglu, G., Ozalp, V. C., Yilmaz, M., Guler, U., Salih, B., Arica, M. Y. (2015). Lysozyme specific aptamer immobilized MCM-41 silicate for single-step purification and quartz crystal microbalance (QCM)-based determination of lysozyme from chicken egg white. Microporous and Meso-porous Materials, 207, 95–104. DOI: 10.1016/j.micromeso.2015.01.009.
Buttry, D. A., Ward, M. D. (1992). Measurement of interfacial processes at electrode surfaces with the electrochemical quartz crystal microbalance. Chemical Reviews, 92, 1355–1379. DOI: 10.1021/cr00014a006.
Chen, Q., Tang, W., Wang, D., Wu, X., Li, N., Liu, F. (2010). Amplified QCM-D biosensor for protein based on aptamer-functionalized gold nanoparticles. Biosensors and Bioelec-tronics, 26, 575–579. DOI: 10.1016/j.bios.2010.07.034.
Cheng, C. I., Chang, Y. P., Chu, Y. H. (2012). Biomolecular interactions and tools for their recognition: focus on the quartz crystal microbalance and its diverse surface chemistries and applications. Chemical Society Reviews, 41, 1947–1971. DOI: 10.1039/c1cs15168a.
Dultsev, F. N., & Tronin, A. V. (2015). Rapid sensing of hepatitis B virus using QCM in the thickness shear mode. Sensors and Actuators B: Chemical, 216, 1–5. DOI: 10.1016/j.snb.2015.04.027.
Gao, C., Yan, D. (2004). Hyperbranched polymers: from synthesis to applications. Progress in Polymer Science, 29, 183–275. DOI: 10.1016/j.progpolymsci.2003.12.002.
Ge, J., Yan, M., Lu, D., Zhang, M., Liu, Z. (2007). Hyper-branched polymer conjugated lipase with enhanced activity and stability. Biochemical Engineering Journal, 36, 93–99. DOI: 10.1016/j.bej.2007.02.018.
Jaruwongrungsee, K., Waiwijit, U., Wisitsoraat, A., Sang-worasil, M., Pintavirooj, C., Tuantranont, A. (2015). Realtime multianalyte biosensors based on interference-free multichannel monolithic quartz crystal microbalance. Biosensors and Bioelectronics, 67, 576–581. DOI: 10.1016/j.bios.2014. 09.047.
Jeong, B., Akter, R., Han, O. H., Rhee, C. K., & Rahman, M. A. (2013). Increased electrocatalyzed performance through dendrimer-encapsulated gold nanoparticles and carbon nanotube-assisted multiple bienzymatic labels: highly sensitive electrochemical immunosensor for protein detection. Analytical Chemistry, 85, 1784–1791. DOI: 10.1021/ac303142e.
Ji, L., Guo, Z., Yan, T., Ma, H., Du, B., Li, Y., Wei, Q. (2015). Ultrasensitive sandwich-type electrochemical immunosensor based on a novel signal amplification strategy using highly loaded palladium nanoparticles/carbon decorated magnetic microspheres as signal labels. Biosensors and Bioelectronics, 68, 757–762. DOI: 10.1016/j.bios.2015.02.010.
Luppa, P. B., Sokoll, L. J., Chan, D. W. (2001). Immuno-sensors—principles and applications to clinical chemistry. Clinica Chimica Acta, 314, 1–26. DOI: 10.1016/s0009-8981 (01)00629-5.
Mao, X., Yang, L., Su, X. L., Li, Y. (2006). A nanoparticle amplification based quartz crystal microbalance DNA sensor for detection of Escherichia coli O157:H7. Biosensors and Bioelectronics, 21, 1178–1185. DOI: 10.1016/j.bios.2005.04. 021.
O’Sullivan, C. K., & Guilbault, G. G. (1999). Commercial quartz crystal microbalances–theory and applications. Biosensors and Bioelectronics, 14, 663–670. DOI: 10.1016/s0956-5663(99)00040-8.
Pei, X., Zhang, B., Tang, J., Liu, B., Lai, W., Tang, D. (2013). Sandwich-type immunosensors and immunoassays exploiting nanostructure labels: A review. Analytica Chimica Acta, 758, 1–18. DOI: 10.1016/j.aca.2012.10.060.
Qian, J., Zhang, C., Cao, X., Liu, S. (2010). Versatile immunosensor using a quantum dot coated silica nanosphere as a label for signal amplification. Analytical Chemistry, 82, 6422–6429. DOI: 10.1021/ac100558t.
Raj, G., Swalus, C., Delcroix, M., Devillers, M., Dupont-Gillain, C., & Gaigneaux, E. M. (2015). In situ quartz crystal microbalance monitoring of the adsorption of poly-oxometalate on a polyampholyte polymer matrix. Journal of Colloid and Interface Science, 445, 24–30. DOI: 10.1016/j.jcis.2014.12.035.
Shen, G., Liu, M., Cai, X., Lu, J. (2008). A novel piezoelectric quartz crystal immnuosensor based on hyperbranched polymer films for the detection of a-fetoprotein. Analytica Chimica Acta, 630, 75–81. DOI: 10.1016/j.aca.2008.09.053.
Shen, G., Cai, C., Wang, K., Lu, J. (2011a). Improvement of antibody immobilization using hyperbranched polymer and protein A. Analytical Biochemistry, 409, 22–27. DOI: 10.1016/j.ab.2010.09.028.
Shen, Z. Q., Wang, J. F., Qiu, Z. G., Jin, M., Wang, X. W., Chen, Z. L., Li, J. W., Cao, F. H. (2011b). QCM immunosensor detection of Escherichia coli O157:H7 based on beacon immunomagnetic nanoparticles and catalytic growth of colloidal gold. Biosensors and Bioelectronics, 26, 3376–3381. DOI: 10.1016/j.bios.2010.12.035.
Tang, D. Q., Zhang, D. J., Tang, D. Y., Ai, H. (2006). Amplification of the antigen-antibody interaction from quartz crystal microbalance immunosensors via back-filling immobilization of nanogold on biorecognition surface. Journal of Immunological Methods, 316, 144–152. DOI: 10.1016/j.jim. 2006.08.012.
Wu, D., Guo, Z., Liu, Y., Guo, A., Lou, W., Fan, D., Wei, Q. (2015). Sandwich-type electrochemical immunosensor using dumbbell-like nanoparticles for the determination of gastric cancer biomarker CA72-4. Talanta, 134, 305–309. DOI: 10.1016/j.talanta.2014.11.025.
Xin, T. B., Chen, H., Lin, Z., Liang, S. X., Lin, J. M. (2010). A secondary antibody format chemiluminescence immunoassay for the determination of estradiol in human serum. Talanta, 82, 1472–1477. DOI: 10.1016/j.talanta.2010.07.023.
Yang, V. C. M., Ngo, T. T. (2000). Biosensors and their applications. New Yourk, NY, USA: Springer.
Yang, F., Yang, Z., Zhuo, Y., Chai, Y., Yuan, R. (2015). Ultrasensitive electrochemical immunosensor for carbohydrate antigen 19-9 using Au/porous graphene nanocomposites as platform and Au@Pd core/shell bimetallic functionalized graphene nanocomposites as signal enhancers. Biosensors and Bioelectronics, 66, 356–362. DOI: 10.1016/j.bios.2014. 10.066.
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Zhang, LQ., Shen, HX., Cheng, Q. et al. Sensitive electrogravimetric immunoassay of hepatitis B surface antigen through hyperbranched polymer bridge linked to multiple secondary antibodies. Chem. Pap. 70, 1031–1038 (2016). https://doi.org/10.1515/chempap-2016-0039
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DOI: https://doi.org/10.1515/chempap-2016-0039