ANP  Vol.2 No.2 , May 2013
Development of Non-Labeled QCM Biosensor for the Detection of β-Galactosidase: A Comparative Study of Gold and Polystyrene Nanoparticles
ABSTRACT

The performance of gold and polystyrene nanoparticles was investigated using quartz crystal microbalance (QCM) as sensor platform; β-Galactosidase antibody with corresponding antigen was utilized for the immunoreaction. The development of the immunosensor included: 1) formation of self assembled monolayers on quartz crystals; 2a) immobilization of p-aminothiophenol functionalized gold nanoparticles on carboxyl-terminated self assembled monolayer, or 2b) immobilization of polystyrene nanoparticles on crystals modified with p-aminothiophenol self assembled monolayer; 3) attachment of monoclonal anti β-Gal on nanoparticles; and 4) detection of target analyte. The nanoparticles used were synthesized in house and characterized by transmission electron microscopy and infrared spectroscopy. The results revealed that antibodies were strongly attached to functionalized gold nanoparticles; the weaker immobilization of antibodies to polystyrene nanoparticles provoked their detachment during antigen detection. When cross reactivity of polystyrene nanoparticles was checked using a different antigen (Brucella), displacement of antibody was not recorded, demonstrating specificity of the reaction. To the best of our knowledge this is the first direct comparison between behaviors of biosensors developed with two commonly used nanoparticles. The results showed that both nanoparticles produced biosensors capable to detect β-Gal. Nevertheless biosensors developed using polystyrene nanoparticles are simpler, cheaper and more eco-friendly than those developed using gold nanoparticles.


Cite this paper
Singh, K. , Choudhary, M. , Chianella, I. and Singh, P. (2013) Development of Non-Labeled QCM Biosensor for the Detection of β-Galactosidase: A Comparative Study of Gold and Polystyrene Nanoparticles. Advances in Nanoparticles, 2, 182-190. doi: 10.4236/anp.2013.22027.
References
[1]   A. V. Fowler and I. Zabin, “The Amino Acid Sequence of Beta Galactosidase. I. Isolation and Composition of Tryptic Peptides,” Journal of Biological Chemistry, Vol. 245, No. 19, 1970, pp. 5032-5041.

[2]   B. W. Matthews, “The Structure of E. coli β-Galactosidase,” C. R. Biologies. Vol. 328, No. 6, 2005, pp. 549 556.

[3]   L. V. Lukacheva, A. A. Zakemovskaya, E. E. Karyakina, I. N. Zorov, A. P. Sinitsyn, M. V. Sukhacheva, et al., “Determination of Glucose and Lactose in Food Products with the Use of Biosensors Based on Berlin Blue,” Journal of Analytical Chemistry, Vol. 62, No. 4, 2007, pp. 388-393.

[4]   M. Porsch-?zcürümez, N. Kischel, H. Priebe, W. Splettst?sser, E. Finke and R. Grunow, “Comparison of Enzyme-Linked Immunosorbent Assay, Western Blotting, Microagglutination, Indirect Immunofluorescence Assay, and Flow Cytometry for Serological Diagnosis of Tularemia,” Clinical and Diagnostic Laboratory Immunology, Vol. 11, No. 6, 2004, pp. 1008-1015.

[5]   B. Koenig and M. Graetzel, “A Novel Immunosensor for Herpes Viruses,” Analalytical Chemistry, Vol. 66, No. 3, 1994, pp. 341-344.

[6]   G. Sauerbrey, “Use of Quartz Crystal Vibrator for Weighting Thin Films on a Microbalance,” Zeitschrift für Physik, Vol. 155, No. 2, 1959, pp. 206-222.

[7]   K. K. Kanazawa and J. Gordon II, “The Oscillation Frequency of a quartz Resonator in Contact with Liquid,” Analytica Chimica Acta, Vol. 175, No. 1, 1985, pp. 99-105.

[8]   D. Kyprianou, A. R. Guerreiro, I. Chianella, E. V. Piletska, S. A. Fowler, K. Karim, et al., “New Reactive Polymer for Protein Immobilisation on Sensor Surfaces,” Biosensors and Bioelectronics, Vol. 24, No. 5, 2009, pp. 1365-1371.

[9]   S. F. Chen, L. Y. Liu, J. Zhou and S. Y. Jiang, “Controlling Antibody Orientation on Charged Self-Assembled Monolayers,” Langmuir, Vol. 19, No. 7, 2003, pp. 2859-2864.

[10]   O. Lazcka, F. J. D. Campob and F. X. M?noz, “Pathogen Detection: A Perspective of Traditional Methods and Biosensors,” Biosensors and Bioelectronics, Vol. 22, No. 7, 2007, pp. 1205-1217.

[11]   D. Kyprianou, A. R. Guerreiro, M. Nirschl, I. Chianella, S. Subrahmanyam, A. P. F. Turner, et al., “The Application of Polythiol Molecules for Protein Immobilisation on Sensor Surfaces,” Biosensors and Bioelectronics, Vol. 25, No. 5, 2010, pp. 1049-1055.

[12]   I. Mannelli, M. Minunni, S. Tombelli and M. Mascini, “Quartz Crystal Microbalance (QCM) Affinity Biosensor for Genetically Modified Organisms (GMOs) Detection,” Biosensors and Bioelectronics, Vol. 18, No. 2, 2003, pp. 129-140.

[13]   M. Pohanka and P. Skladal, “Piezoelectric Immunosensor for the Direct and Rapid Detection of Francisella tularensis,” Folia Microbiologica, Vol. 52, No. 4, 2007, pp. 325-330.

[14]   X. Mao, L. Yang, Su X Li and Y. Li, “A Nanoparticle Amplification Based Quartz Crystal Microbalance DNA Sensor for Detection of Escherichia Coli O157:H7,” Biosensors and Bioelectronics, Vol. 21, No. 7, 2006, pp. 1178-1185.

[15]   M. K. Choudhary, R. Agrawal, R. Kumar, P. Singh, B. R. K. Gupta and K. P. Singh, “Detection of cadmium-re sistant Rhizobacteria Using Piezoelectric Nanobiosensor,” International Journal of Nanoscience, Vol. 9, No. 5, 2010, pp. 461-469.

[16]   R. Colorado Jr., R. J. Villazana and T. R. Lee, “Self Assembled Monolayers on Gold Generated from Aliphatic Dithiocarboxylic Acids,” Langmuir, Vol. 14, No. 22, 1998, pp. 6337-6340.

[17]   A. Tlilli, A. Abdelghani, S. Hleli and M. A. Aaaref, “Electrical Characterization of a Thiol SAM on the Gold as a First Step for the Fabrication of Immunosensors Based on a Quartz Crystal Microbalance,” Sensors, Vol. 4, No. 6, 2004, pp. 105-114.

[18]   I. Lee, X. Wang, C. F. Zhu, C. Wang and C. Bai, “Investigation of Polystyrene Nanoparticles and DNA-Protein Complexes by AFM with Image Reconstruction,” Applied Surface Science, Vol. 126, No. 3-4, 1998, pp. 281-286.

[19]   K. K. Kar and P. Parik, “High Molecular Weight Polystyrene Nanospheres,” 2005.

[20]   E. J. C. Kellar, C. Galiotis and E. H. Andrews, “Raman Vibrational Studies of Syndiotactic Polystyrene. 1. Assignments in a Conformational/Crystallinity Sensitive Spe ctral Region,” Macromolecules, Vol. 29, No. 10, 1996, pp. 3515-3520.

[21]   Z. Li, J. Zhou, Z. Zhang and H. Dang, “Self-Assembly of Carboxyl Functionalized Polystyrene Nano-Spheres into Close-Packed Monolayers via Chemical Adsorption,” Chinese Journal of Chemistry, Vol. 22, No. 10, 2004, pp. 1133-1137.

[22]   V. V. Shmanai, T. A. Nikolayeva, L. G. Vinokurova and A. A. Litoshka, “Oriented Antibody Immobilization to Polystyrene Macrocarriers for Immunoassay Modified with Hydrazide Derivatives of Poly(meth)acrylic Acid,” BMC Biotechnology, Vol. 1, No. 1, 2001, p. 4. http://www.biomedcentral.com/1472-6750/1/4

[23]   W. Qian, D. Yao, F. Yu, B. Xu, R. Zhou, X. Bao and Z. Lu, “Immobilization of Antibodies on Ultraflat Polystyrene Surfaces,” Clinical Chemistry, Vol. 46, No. 9, 2000, pp. 1456-1463.

 
 
Top