Highly sensitive potentiometric immunosensor for hepatitis B surface antigen diagnosis
Tóm tắt
A highly sensitive potentiometric immunosensor for the diagnoses of epidemic diseases has been developed by means of self-assembly to immobilize hepatitis B surface antibody (HBsAb) for the detection of hepatitis B surface antigen (HBsAg) as a model. At first, the Nafion containing ™SO
3
™
groups was immobilized on a platinum electrode surface to absorb the ™NH3
+ groups of antibody moleculesvia the opposite-charged adsorption technique, in the meantime, hepatitis B surface antibodies were adsorbed onto the surface of Au nanoparticles, then hepatitis B surface antibodies and Au nanopartilces were entrapped into polyvinyl butyral on the surface of Nafion film. The modified procedure was further characterized by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). The influence and factors influencing the performance of resulting immunosensor were studied in detail. The resulting immunosensor exhibited sigmoid curve with log HBsAg concentrations, high sensitivity, wide linear range from 26 to 1280 ng · mL−1 with a detection limit of 3.1 ng · mL−1, rapid potentiometric response (σ min) and long-term stability (/s>4 months). Analytical results of clinical samples show that the developed immunoassay is comparable with the enzyme-linked immunosorbent assays (ELISAs) method, implying a promising alternative approach for detecting HBsAg in the clinical diagnosis.
Tài liệu tham khảo
Bunde, R. L, Jarvi, E. J., Rosentreter, J. J., Piezoelectric quartz crystal biosensors, Talanta, 1998, 46: 1223–1236.
Wang, H., Li, J. H., Ding, Y. J. et al., Novel immunoassay for Toxoplasma gondii-specific immunoglobulin G using a silica nanoparticle-based biomolecular immobilization method, Analytica Chimica Acta, 2004, 501: 37–43.
Wang, H., Lei, C. X., Li, J. S. et al., A piezoelectric immunoagglutination assay for Toxoplasma gonlii antibodies using gold nanoparticles, Biosensors and Bioelectronics, 2004, 19: 701–709.
Wang, H., Dan, L., Wu, Z. Y. et al., A reusable piezo-immuno-sensor with amplified sensitivity for ceruloplasmin based on plasma-polymerized film. Talanta, 2004, 62: 201–208.
Liu, G. D., Wu, Z, Y, Wang, S. P. et al., Renewable amperometric immunosensor for Schistosoma japonium antibody assay. Anal. Chem., 2001, 73: 3219–3226.
Zhang, S. S., Jiao, K., Chen, H. Y. et al., Detection of ferritin in human serum with a MAP-H2O2-HRP voltammetric enzymelinked immunoassay system. Talanta, 1999, 50(1): 95–101.
Du, D., Yan, F., Liu, S. L. et al., Immunological assay for carbohydrate antigen 19-9 using an electrochemical immunosensor and antigen immobilization in titania sol-gel matrix, J. Immuno. Methods, 2003, 283: 67–75.
Ye, Y. K., Zhao, J. H., Yan, F. et al., Electrochemical behavior and detection of hepatitis B virus DNA PCR production at gold electrode, Biosens. Bioelectron., 2003, 18: 1501–1508.
Yan, G. F., Ju, H. X., Liang, Z. C. et al., Technical and clinical comparison of two fully automated methods of CA 125 immunoassay in serum, J. Immuno. Methods, 1999, 225: 1–8.
Chen, H. Y., Ju, H. X., Xun, Y. G., Methylene blue/perfluorosul-fonated ionomer modified microcylinder carbon fiber electrode and its application for the determination of hemoglobin, Anal. Chem., 1994, 66: 4538–4542.
Liang, R. P., Qiu, J. D., Zou, X. Y. et al., Preparation and application of sol-gel-HBsAb immunosensor, Chemical J. Chinese Universities, 2003, 5: 793–797.
Liang, R. P., Qiu, J. D., Zou, X. Y. et al., The self-assembled HBsAb monolayer gold electrode and its application to HBsAg determination, Appl. Chem. (in Chinese), 2001, 18: 705–708.
Frens, G., Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions [J], Nature Phy. Sci., 1973, 241:20–22.
Li, L. D., Li, W. J., Sun, C. Q., Fabrication of chemically-modified electrode containging 2: 18 tungstophosphate anions based on sol-gel technique and its electrocatalysis, Chemical J. Chinese Universities, 2000, 21(6): 865–869.
Jiang, H. H., Zou, H. F., Wang, H. L. et al., Combination of MALDI-TOF mass spectrometry with immobilized enzyme microreactor for peptide mapping, Science in China, Ser. B, 2000, 43(6): 625–633.
Li, M. L., Li, B. F., Jiang, L. et al., Two orientations and photochromism of polyelectrolyte/bacteriorhodopsin alternative deposition multilayers, Science in China, Ser. B, 2000, 43(3): 313–322.