Microarrays preparing analytes

Arenkov et al. prepared poly(acrylamide) gel pads for use in protein microarrays [199], The gels were prepared by photopolymerization of acrylamide and crosslinkers. Capture probes were immobilized, either by use of glutaraldehyde or by converting some of the acrylamide groups into hydrazides and subsequent coupling of aldehyde-modified antibodies to the pending hydrazide groups. Then, immunoassays were performed on the pads, either assays with directly labeled analytes or sandwich assays. Furthermore, the gel pads were used for enzyme activity studies. 

Other extensions that have been made include the construction of a biochip in which a reverse transcription PCRprocess can be carried out [384] and the integration of PCR with capillary electrophoresis [385], DNA microarray hybridization [386] and sample preparation [387,388]. The speed of analysis, the ease of integrating different functions and the relative low costs of micro-PCR production are important advantages of the miniaturized PCR technique that suggest that this bioorganic microreactor device will be thoroughly implemented in many analytical labs. 

Recently, a small molecule fluorophore phosphosensor technology referred as Pro-Q Diamond dye has been developed to detect and quantitate phosphorylated amino acids within peptides and proteins in microarrays. ° In addition to binding assays, fluorescence detection methods have also been developed for functional assays. For example, microarrays of quenched fluorescent substrates can be used to detect protease or esterase activities in the analytes. In this method, quenched fluorescent substrates are prepared by coupling the peptide substrate to coumarin, a fluorescent dye. These peptide substrates are then spotted onto the solid support... 

The electrochemical biosensor method has shown high potential in microarray usage for its sensitivity, reproducibility, selectivity, and reversibility. Unlike other methods, the biosensor method depends on the electrochemical property of the coating receptors and binding analytes. It is prepared by electrochemical synthesis of polymers in situ on platinum electrodes. This technology, however, will require further... 

Rogers Y H, Jiang-Baucom P, Huang Z J, et al. (1999). Immobilization of oligonucleotides onto a glass support via disulfide bonds A method for preparation of DNA microarrays. Analyt. Biochem. 266 23-30. 

Overall, these results demonstrated that the binding of lectins with the prepared carbohydrate microarrays are specific and multiple-analytic characterization can be achieved on the aminooxyacetyl-functionalized slide with good selectivity. Furthermore, periodate oxidation of the immobilized oligosaccharides with NaI04 resulted in the loss of lectin binding. These experiments verify that the fabricated carbohydrate microarray is well suited for the selective identification of carbohydrate-binding proteins. 

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