Antibody chips

Fig. 4 Antibody chip technology. Chips with immobilized antibodies are probed with a specimen and assessed for binding of specific disease marker proteins...

Endo, T., Okuyama, A., Matsubara, Y., Kobayashi, M., Morita, Y., Mizukami, H., Tamiya, E., Monitoring of coplanar polychlorinated biphenyls (Co-PCB) by the multi flow antibody chip. Micro Total Analysis Systems 2003, Proceedings 7th jlTAS Symposium, Squaw Valley, CA, Oct. 5-9, 2003, 567-570. 

Antibody chips. These consist of arrayed antibodies and are used to detect and quantify specific proteins in a complex mixture. They can be thought of as miniaturized high-throughput immvmoassay devices. 

Ogasawara, D., Hirano, Y, Yasukawa, T., Shiku, H., Kobori, K., Ushizawa, K., Kawabata, S., Matsue, T. Electrochemical microdevice with separable electrode and antibody chips for simultaneous detection of pepsinogens 1 and 2. Biosens Bioelectmn 2006, 21, 1784-1790. 

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... 

The ProteinChip System from Ciphergen Biosystems uses patented SELDI (Surface-Enhanced Laser Desorption/Ionization) ProteinChip technology to rapidly perform the separation, detection, and analysis of proteins at the femtomole level directly from biological samples. ProteinChip Systems use ProteinChip Arrays which contain chemically (cationic, anionic, hydrophobic, hydrophilic, etc.) or biochemically (antibody, receptor, DNA, etc.) treated surfaces for specific interaction with proteins of interest. Selected washes create on-chip, high-resolution protein maps. This protein mass profile, or reten-tate map of the proteins bound to each of the ProteinChip Array surfaces, is quantitatively detected in minutes by the ProteinChip Reader. 

In a second experiment, Cy5-labelled antiBSA antibodies were immobilised on a silanised glass slide precoated with metallic nanoislands using a polydimethylsiloxane (PDMS) flow-cell. The antibody solution was left for 1 hour to attach and then the cell was flushed with deionised water. The slide was then dried with N2. For this experiment, a portion of the slide was not coated with metallic nanoislands, in order to act as a reference. Figure 20 shows the image recorded using the fluorescence laser scanner mentioned previously. The enhancement in fluorescence emission between those areas with and without nanoislands (B and A, respectively) is again evident. For both chips, an enhancement factor of approximately 8 was recorded. There is considerable interest in the elucidation and exploitation of plasmonic effects for fluorescence-based biosensors and other applications. 

Biochips can be used for either measuring differential expression between two populations or for testing for the presence of a DNA sequence (resequencing). Protein chips have been applied in expression profiling and antibody detection, binding specificities of a protein expression library and protein-protein interactions. 

Monoclonal and polyclonal antibodies serve as specific affinity ligands in protein chips. Though monoclonal/polyclonal pairs are more readily available than monoclonal/monoclonal pairs, polyclonal antibodies often cause higher background and lower sensitivity and specificity. 

Duan, L., Wang, Y., Li, S.S-C., Wan, Z., and Zhai, J. (2005) Rapid and simultaneous detection of human hepatitis B virus and hepatitis C virus antibodies based on a protein chip assay using nano-gold immunological amplification and silver staining method. BMC Infect. Dis. 5, 53. http //www.biomedcentral. com/1471-2334/5/53. 

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