Peptide microarrays

Salisbury CM, Maly DJ, Ellman JA (2002) Peptide microarrays for the determination of protease substrate specificity. J Am Chem Soc 124 14868-14870... 

Andresen H, Bier FF (2009) Peptide microarrays for serum antibody diagnostics. Methods Mol Biol 509 123-134... 

Diluted human serum (1 pL) was incubated with the peptide microarray and bound antibodies were detected using a rhodamine-labeled anti-human IgG. Signal was detected using a slide scanner (Affymetrix model 418) with data collection in the Cy3 channel. A reported eightfold gain in sensitivify at 100% specificity over standard ELISA was achieved using the peptide microarray. 

Key Words Peptide microarrays small-molecule microarrays small-molecule immobilization peptide immobilization chemoselective ligation. 

Oligonucleotide and peptide microarrays can be prepared in situ with light-directed synthesis on a glass surface in conjunction with either a photolithographic method or using a micromirror device (2,11-15). However, these methods are not useful for most organic synthesis. Furthermore, such approaches require equipment that is not readily available. Spot synthesis on cellulose membrane is another in situ synthesis method, but the resulting microarrays are low density (16). 

Inoue Y, Mori T, Yamanouchi G, Han X, Sonoda T, Niidome T, Katayama Y (2008) Surface plasmon resonance imaging measurements of caspase reactions on peptide microarrays. Anal Biochem 375 147-149... 

Protein Microan ay Peptide Microarray Antibody Microarray... 

Houseman et al. prepared a peptide chip by Diels-Alder-mediated reaction of kinase-peptide substrates with a self-assembled monolayer of alkanethiolates on gold surface. Peptide phosphorylation was determined by incubating the peptide microarrays with c-src protein-tyrosine kinase followed by quantitation with a phos-phoryl imager. In the presence of soluble inhibitors at a range of concentrations, dose-dependent inhibition of phosphorylation against a number of peptide substrates could be determined on a single chip. ... 

Peptide microarrays are prepared by immobilizing many peptide molecules on the surface of a solid support in a small area in an addressable fashion. The immobilization can be achieved via in situ synthesis or chemical ligation through a covalent bond. A hydrophihc linker between the sohd surface and the peptide usually is added to minimize steric hinderance caused by the sohd support. The most commonly used solid support for microarray printing is a standard microscope glass slide. Other solid supports also have been used such as polystyrene, nitrocellulose membranes, PVDF membranes, Hybond ECL membranes, gold surfaces, and chemical vapor deposited diamond films. 

We reported site-specific ligation of peptides to a carrier protein or polymer before printing onto the glass slide with an automatic arrayer to form a peptide microarray (22). Recently, Dikmans et al. (23) reported a novel process for manufacturing... 

Prepare peptide microarray using digital chemistry http //gaolab.chem. uh.edu/resl.htm... 

Chemical diversity for chemical microarrays can be accessed in two ways importing compounds onto the chip surface or synthetically creating molecules directly on the support. In the pioneering work by Fodor and coworkers, a combination of solid-phase organic synthesis and photolithographic techniques was applied to the in siiu creation of oligonucleotide and peptide microarrays [54, 55]. 

Gosalia, D. N., Salisbury, C. M., Ellman, J. A., and Diamond, S. L. 2005. High throughput substrate specificity profiling of serine and cysteine proteases using solution-phase fluorogenic peptide microarrays. Mol. Cell. Proteomics 4 626-36. 

Advantages of the on-chip approaches are that the systems are well established and very uniform processing is practicable. But these approaches can only be applied for the production of oligonucleotide microarrays. So far, only scientific but no commercial approaches have been undertaken to produce peptide microarrays by an on-chip approach. Additionally, depending on the specific application, low-density microarrays and only low microarray numbers are sufficient to answer particular biological questions thus, often small batches (hundreds) of microarrays with low density (a few hundred spots) are needed. In research applications the array layout needs to be flexible. The production of protein or cell microarrays is in high demand as well. 

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