Implementation Of Protein Microarrays

Implementation of protein microarray strategies will help to address the different features of proteins that may be altered in disease. These include biochips that contain either peptides or proteins as well as recombinant proteins that can be synthesized directly onto the chip [28]. The development of protein microarrays relies on the use of capture agents, such as ribozymes, partial molecules, antibodies, and modified binding proteins (Fig. 5.3). Newer technologies have been developed to immobilize the entire protein repertoire of tissue using a reverse-phase

FIGURE 5.3 Antibody microarray example. Approximately 2500-3500 cells of each of six histologic types were microdissected and biotinylated protein from each dissection was used for a single incubation on a single antibody array. Examples of P-SCAN analysis indicated that Stat5a, EGFR-nonphosphoY1173, PR, and Rsk proteins decreased in expression in epithelium with advancing epithelial disease. Lines and circles result from the orientation function and data analysis by P-SCAN. (Proteomics, Knezevic et al. (2001). Copyright John Wiley and Sons Ltd. Reproduced with permission.) See Plate 5.3 in Color Plate Section.

FIGURE 5.3 Antibody microarray example. Approximately 2500-3500 cells of each of six histologic types were microdissected and biotinylated protein from each dissection was used for a single incubation on a single antibody array. Examples of P-SCAN analysis indicated that Stat5a, EGFR-nonphosphoY1173, PR, and Rsk proteins decreased in expression in epithelium with advancing epithelial disease. Lines and circles result from the orientation function and data analysis by P-SCAN. (Proteomics, Knezevic et al. (2001). Copyright John Wiley and Sons Ltd. Reproduced with permission.) See Plate 5.3 in Color Plate Section.

protein array [29]. Using this approach, pro-survival checkpoints were determined showing the transition for histologically normal to neoplastic prostate tissue associated with a change in phosphorylation status of signal subpopulations. Similarly, protein microarrays can be incubated with patient serum and used to detect autoantibody binding to specific proteins of autoimmune disease, such as systemic lupus erythe-matosus and rheumatoid arthritis [30]. This new area of research is limited by the availability of the capture proteins and antibodies.

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