Antibody sensor

Y.-M. Zhou, Z.-Y. Wu, G.-L. Shen, and R.-Q. Yu, An amperometric immunosensor based on Nafion-modified electrode for the determination of Schistosoma japonicum antibody. Sensors and Actuators, B Chemical 89, 292-298 (2003). 

MIP-based assay and the MIP sensor chip was able to be regenerated many more times than the antibody sensor chip. In another application reviewed by Nedelkov and Nelson [ 173], the detection and quantitation abilities of SPR technology were coupled with mass spectral identification of proteins captured, either directly on the sensor chip or following elution from the chip surface. 

FIGURE 6.9 Basic principle of a fiber optic antigen-antibody sensor. (Taken from Anderson G.P., Golden J.P., and Ligler F.S. 1993. IEEE Trans. Biomed. Eng. 41 578.)... 

Matsushita M, Yoshida K, Yamamoto N et al (2003) High-throughput screening by using a blue-fluorescent antibody sensor. Angew Chem Int Ed 42 5984—5987... 

Sun X, Du S, Wang X, Zhao W, Li Q (2011) A label-free electrochemical immunoscmsor for carbofuran detection based on a sol-gel entrapped antibody. Sensors 11 9520-9531... 

Enzyme Immunosensors. Enzyme immunosensors are enzyme immunoassays coupled with electrochemical sensors. These sensors (qv) require multiple steps for analyte determination, and either sandwich assays or competitive binding assays maybe used. Both of these assays use antibodies for the analyte of interest attached to a membrane on the surface of an electrochemical sensor. In the sandwich assay type, the membrane-bound antibody binds the sample antigen, which in turn binds another antibody that is enzyme-labeled. This immunosensor is then placed in a solution containing the substrate for the labeling enzyme and the rate of product formation is measured electrochemically. The rate of the reaction is proportional to the amount of bound enzyme and thus to the amount of the analyte antigen. The sandwich assay can be used only with antigens capable of binding two different antibodies simultaneously (53). 

Biosensors ai e widely used to the detection of hazardous contaminants in foodstuffs, soil and fresh waters. Due to high sensitivity, simple design, low cost and real-time measurement mode biosensors ai e considered as an alternative to conventional analytical techniques, e.g. GC or HPLC. Although the sensitivity and selectivity of contaminant detection is mainly determined by a biological component, i.e. enzyme or antibodies, the biosensor performance can be efficiently controlled by the optimization of its assembly and working conditions. In this report, the prospects to the improvement of pesticide detection with cholinesterase sensors based on modified screen-printed electrodes are summarized. The following opportunities for the controlled improvement of analytical characteristics of anticholinesterase pesticides ai e discussed ... 

Bulk and surface imprinting strategies are straightforward tools to generate artificial antibodies. Combined with transducers such as QCM (quartz crystal microbalance), SAW (surface acoustic wave resonator), IDC (interdigital capacitor) or SPR (surface plasmon resonator) they yield powerful chemical sensors for a very broad range of analytes. 

The aim of our investigation was the development of the amperometric enzyme immunosensor for the determination of Klebsiella pneumoniae bacterial antigen (Ag), causes the different inflammatory diseases. The biosensing pail of the sensors consisted of the enzyme (cholinesterase) and antibodies (Ab) immobilized on the working surface of the screen-printed electrode. Bovine seiaim albumin was used as a matrix component. 

The working conditions of the immunosensor (enzyme and antigen concentrations, dilutions of the antibodies, pH of the buffer solution) were found. The cholinesterase immobilized demonstrated the maximum catalytic activity in phosphate buffer solution with pH 8.0. The analytical chai acteristics of the sensor - the interval of the working concentrations and detection limit - have been obtained. The proposed approach of immunoassay made possible to detect 5T0 mg/ml of the bacterial antigen. 

Sandwich-type sensors are applicable for measuring large antigens that are capable of binding two different antibodies. Such sensors utilize an antibody that binds the analyte-antigen, which then binds an enzyme-labeled second antibody. After removal of the nonspecifically adsorbed label, the probe is placed into the substrate-containing solution, and the extent of the enzymatic reaction is monitored... 

Other sensor applications can be considered if some sensitive biological molecules (such as antibodies or receptors) are attached to the nanogranule. If, for example, an antibody molecule is attached to it, then the granule is placed between two electrodes, and single-electron current flows between them. The step value of the coulomb staircase depends on the capacity of the junctions. When the antibody molecule binds specific antigen, the capacity value will be changed, and, therefore, the step value of the VH characteristics will also change. 

Fig.Sa-f. The sensorgram of the repeated injection of the aqueous viologen dimer 2 (a, c, e) and the antibody (b, d, f) solutions. [Viologen dimer 2]=2.0 pM and [antibody]=2.0 pM in phosphate borate buffer. Injection period 60 s for a-c and 120 s for d-f. A solution of viologen dimer 2 or the antibody passes over the surface of the sensor chip for 60 or 120 s at a constant flow rate of 20 pL min. The surface of the sensor chip was subsequently washed with buffer...

The amount of antibody immobilized on the sensor chip decreased with increasing concentration of methyl viologen. To enlarge the difference in the sig-... 

Fig. 7. A proposed structure of the complex of the antibody with the trivalent antigen immobilized on the surface of the sensor chip...

Fig. 12a,b. The sensorgrams for the binding of the antibody dendrimer (a) or IgG (b) to the anionic porphyrin immobilized onto the surface of the sensor chip. Phosphate borate buffer (0.1 M, pH 9.0) was used. TCPP was immobilized via hexamethylenediamine spacer onto the sensor chip and then a solution of IgG or the dendrimer was injected to the flow cell. After 60 s from the injection of the antibody solutions, flow ceU was filled with buffer... 

Fig. 13a-e. The increase of the signal intensities by the addition of the dendritic complexes composed of IgGs and protein A. The hapten was immobilized to the surface of the SPR sensor chip. The increase of the signal intensities on the complex formation of hapten with the antibodies were monitored. The addition of mouse IgG specific for hapten (Abl) (a), the complex of the Abl with protein A (b), one to one complex of Abl with anti-mouse IgG (Fc) antibody (Ab2) (c), two to one complex of Abl with Ab2 (d), and two to one complex of Abl with Ab2 in the presence of protein A (e)... 

An enhancement of SPR signal intensity was observed by the addition of the antibody to the divalent antigen-antibody complex immobilized onto the surface of the sensor chip, indicating the formation of linear supramolecules. An amplification method of the detection signals for a target molecule has been... 

---