Specificity, immunosensor

The development of immunosensors is one of the most active research areas in immun-odiagnostics. A large number of immunosensors, which combine the sensitivity and specificity of immunoassays with physical signal transduction, have been developed... 

A major disadvantage is that the direct sensor detection cannot distinguish between the sensor response to the specific analyte binding from the response to a possible nonspecific adsorption of other compounds. The nonspecific fouling from blood or blood serum seems to be one of the main barriers for practical application of immunosensors in medical diagnostics. 

There is a continuing demand for fast and simple analytical methods for the determination of many clinical, biochemical and environmental analytes. In this respect, immunoassays and immunosensors that rely on antibody-antigen interactions provide a promising means of analysis owing to their specificity and sensitivity. High specificity... 

Due to the broad substrate specificity of AP, and the drive for higher efficiency, several studies have recently investigated the suitability of alternative substrates to the common 4-APP [47, 48], For example, Pemberton et al. compared 4-APP and 1-naphthyl phosphate (1-NP) as AP substrates in an amperometric immunosensor for progesterone [47], The signal generation scheme when 1-NP is used as a substrate is illustrated in Scheme 2. 

C. Singh, G.S. Agarwal, G.P. Rai, L. Singh, and V.K. Rao, Specific detection of Salmonella typhi using renewable amperometric immunosensor. Electroanalysis 17, 2062-2067 (2005). 

Since immunosensors usually measure the signals resulting from the specific immu-noreactions between the analytes and the antibodies or antigens immobilized, it is clear that the immobilization procedures of the antibodies (antigens) on the surfaces of base transducers should play an important role in the construction of immunosensors. Numerous immobilization procedures have been employed for diverse immunosensors, such as electrostatic adsorption, entrapment, cross-linking, and covalent bonding procedures. They may be appropriately divided into two kinds of non-covalent interaction-based and covalent interaction-based immobilization procedures. 

There are mainly three types of transducers used in immunosensors electrochemical, optical, and microgravimetric transducers. The immunosensors may operate either as direct immunosensors or as indirect ones. For direct immunosensors, the transducers directly detect the physical or chemical effects resulting from the immunocomplex formation at the interfaces, with no additional labels used. The direct immunosensors detect the analytes in real time. For indirect immunosensors, one or multiple labeled bio-reagents are commonly used during the detection processes, and the transducers should detect the signals from the labels. These indirect detections used to need several washing and separation steps and are sometimes called immunoassays. Compared with the direct immunosensors, the indirect immunosensors may have higher sensitivity and better ability to defend interference from non-specific adsorption. 

C. Fernandez-Sanchez, C.J. McNeil, K. Rawso, and O. Nilsson, Disposable noncompetitive immunosensor for free and total prostate-specific antigen based on capacitance measurement. Anal. Chem. 76, 5649-5656 (2004). 

In an immunosensor the core-cover interface of an optical waveguide structure is coated with a chemo-optical transducer receptor layer, which can selectively bind to specific analyte molecules present in the cover medium. The receptor-analyte reaction obeys the law of mass action, which states that the rate of a reaction is proportional to the concentration of the reactants. At equilibrium, the rate of formation of the receptor-analyte complex is equal to the rate of breaking, and the equilibrium constant, K. can be written as... 

Immunosensors have been developed commercially mostly for medical purposes but would appear to have considerable potential for food analysis. The Pharmacia company has developed an optical biosensor, which is a fully automated continuous-flow system which exploits the phenomenon of surface plasmon resonance (SPR) to detect and measure biomolecular interactions. The technique has been validated for determination of folic acid and biotin in fortified foods (Indyk, 2000 Bostrom and Lindeberg, 2000), and more recently for vitamin Bi2. This type of technique has great potential for application to a wide range of food additives but its advance will be linked to the availability of specific antibodies or other receptors for the various additives. It should be possible to analyse a whole range of additives by multi-channel continuous flow systems with further miniaturisation. 

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