ABSTRACT
This paper reports a simple, sensitive, and reusable piezoelectric immunosensor using magnetic hydroxyapatite (HAP)/gamma-Fe(2)O(3)/Au nanocomposite. Use of porous HAP nanocrystals embedded with gamma-Fe(2)O(3) and colloidal gold nanoparticles resulted in a multifunctional HAP/gamma-Fe(2)O(3)/Au nanocomposite. Under optimized conditions, the biocompatible nanocomposites were exploited for direct adsorption of large quantities of rabbit anti-human immunoglobulin G antibodies (anti-hIgG) with well-preserved immunoactivity. In a homogeneous bulk solution, the hIgG analytes were captured by the anti-hIgG-derivatized immunocomposites followed by magnetic separation/enrichment onto a bovine serum albumin (BSA)-sealed QCM probe before measuring. This QCM immunosensor can quantitatively determine concentrations of hIgG ranging from approximately 20 to 800 ng/ml, with a detection limit of approximately 15 ng/ml. Moreover, regeneration of the immunosensor can be simply realized by canceling the controllable magnetic field. With the possibility of performing the analysis automatically and considering its ease of use, high sensitivity, and good reusability, this magnetic separation-assisted QCM immunosensor may have great potential to be further tailored as a general and promising alternative for a broad range of practical applications.
Subject(s)
Antibodies/analysis , Biosensing Techniques/instrumentation , Biosensing Techniques/methods , Immunoglobulin G/analysis , Magnetics , Nanoparticles/chemistry , Adsorption , Animals , Antibodies/immunology , Cattle , Colloids/chemistry , Durapatite/chemistry , Durapatite/immunology , Ferric Compounds/chemistry , Ferric Compounds/immunology , Gold/chemistry , Gold/immunology , Humans , Hydrogen-Ion Concentration , Immunoglobulin G/immunology , Particle Size , Sensitivity and Specificity , Serum Albumin, Bovine/chemistry , Serum Albumin, Bovine/immunology , Surface Properties , Swine , Time FactorsABSTRACT
A direct electrochemical biosensing platform has been fabricated by covalent incorporation of carbon nanotubes (CNT) and gold nanoparticles (GNP) onto the poly(thionine) (PTH) film deposited by electropolymerization. With the synergic effects of the composite nanomaterials together with the excellent mediating redox polymer, the proposed platform could allow for faster electron transfer and higher enzyme immobilization efficiency than the platforms designed by using CNT or GNP alone. Comparison studies indicated that the as-developed H(2)O(2) sensor could show greatly improved performances of amperometric responses.
Subject(s)
Biosensing Techniques/methods , Gold/chemistry , Nanotubes, Carbon/chemistry , Phenothiazines/chemistry , Polymers/chemistry , Biosensing Techniques/instrumentation , Electrochemistry , Electrodes , Hydrogen Peroxide/analysis , Nanoparticles , Oxidation-Reduction , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
BACKGROUND: The resurgence of the parasitic disease schistosomiasis calls for more efficient diagnostic tests. We developed a rapid, simple, portable, and sensitive immunoagglutination assay that uses SiO(2) particles and quartz crystal microbalance (QCM) for quantifying Schistosoma japonicum (Sj) antibodies (SjAb). METHODS: We prepared submicrometer-sized silica particles derivatized with Sj antigens as replacements for traditional latex microspheres to specifically agglutinate in the presence of SjAb targets, and we used the QCM monitor to measure the resulting frequency shifts. We optimized the assay medium by adding poly(ethylene glycol) (PEG) as a response accelerator of immunoagglutination. To minimize or eliminate any nonspecific agglutination or adsorption interferences, we conducted appropriate sealing procedures separately for silica particles and the QCM probe. RESULTS: The measured frequency changes were linearly related to the SjAb concentrations in infected rabbit serum. The PEG-assisted immunoagglutination system was quantitatively sensitive to SjAb concentrations ranging from approximately 0.70 to 32.31 mg/L, with a detection limit of approximately 0.46 mg/L. The obtained linear regression equation was: y=43.61 x+80.44 (r=0.9872). Several serum specimens were evaluated with the developed QCM immunoassay and the results were compared with ELISA, validating the feasibility of practical applications. CONCLUSIONS: This novel immunoagglutination-based QCM detection format is rapid, simple to use, and more portable than conventional diagnostic immunoassays, thus offering a promising alternative tool that can be used for point-of-care clinical diagnosis of schistosomiasis, particularly in epidemic situations.
