Many researchers have studied in vivo combinatorial biotechnology, e.g., either phage or cell-surface display techniques, and developed polypeptide sequences, which can specifically bind to metals [5�C7], oxides [8,9], and semiconductors [10�C12]. Among them, gold-binding polypeptide (GBP) is one of the genetically http://www.selleckchem.com/products/Bicalutamide(Casodex).html engineered proteins for a strong binding onto the gold surface [7,13,14]. Whereas many proteins and chemicals bind to the gold surface with thiol linkage, GBP does not contain a cysteine residue having thiol group. Although the definite mechanism is not clear yet, it is estimated that these polar groups in GBPs seem to coordinately interact with the gold surface within a monolayer [15,16]. In addition, the kinetics and thermodynamics of biomimetic interactions between the GBP, and the gold surface were investigated by surface plasmon resonance (SPR) [14,17,18].
Compared with other thiol-based systems, the GBP binds tightly to the gold surface due to the lower standard Gibbs free energy for the bond, and the binding process is fast under aqueous conditions compatible with biological environments [14,17,19]. These characteristics suggest its potential applications in nano- and bio-technologies as novel agents for surface functionalization [13].Moreover, immobilization with correct orientation of biological material is a problem of prime importance in biosensors. We employed GBP-fusion proteins in the construction of biosensor for the detection of hepatitis B viral surface antigen (HBsAg) as a model (Figure 1), which is a biomarker for diagnosing the hepatitis B virus (HBV).
The strong affinity between the GBP and the gold surface guarantees the stability of this sensor system and orients the sensing parts outward from the solid surface, exposing them directly to the target sample [13,16]. Furthermore, electrochemical detection has attracted considerable interest recently for miniaturized analytical systems [20,21], including remarkable sensitivity (approaching that of fluorescence), inherent miniaturization of both the detector and control instrumentation, independence of optical path length or sample turbidity, low cost, low-power requirements and high compatibility [22,23]. Besides, one of the most attractive points of this method is its potential for portable assays in a variety of point-of-care testing (POCT) environments.
We here developed a simple platform biosensor technology by mediating Cilengitide the recognition parts and the solid surface on the gold substrate. SPR analyses were used for optimization of sample concentrations and verification of target sensing. Electrical signal-based selleck Ponatinib detection methods for HBV such as electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) were developed on the gold electrode surface, which has been a very versatile material in the field of biosensors.Figure 1.