Protein-Detecting Microarrays

J. Am. Chem. Soc. 2000, 122, 7849-7850 (b) Printing Small Molecules as Microarrays and Detecting Protein-Ligand Interactions en Masse, G. MacBeath,... 

The protein microarray represents an emerging technology. While we have described its potential utility, several key problems remain to be overcome before this tool is fully adopted by the research and biopharmaceutical commxmities. The most likely first embodiment will be an antibody "protein-detecting" microarray. This is understandable based upon the availability and suitability of antibody libraries originally developed for ELISA. We have discussed many demonstrahons of antibody arrays in this chapter but commercial introductions (Pierce, Beckman Coulter) have been limited. 

MacBeath, G., Koehler, A.N., Schreiber, S.L. (1999) Printing small molecules as microarrays and detecting protein.Kgand interactions en masse./Aw Chem Soc. 121, 7967-7968... 

Fig. 3. Schematic of staining process of SARS-CoV immunochip. (1) Spotting A high-precision robot transfers the samples, SARS-CoV proteins, and glycans of various complexities, from 96-well plate to nitrocellulose-coated glass slides. (2) Staining Before staining, the slides are rinsed with IX phosphate-buffered saline (PBS), and blocked with 1% bovine serum albumin (BSA)-PBS containing 0.05% NaN3 and 0.05% Tween-20. They are subsequently incubated with horse anti-SARS sera. The primary antibodies captured by microarrays are detected using biotinated anti-horse immunoglobulin (Ig)G, and visualized by Cy3-streptavidin.

G. MacBeath, A. N. Koehler, and S. L. Schreiber, Printing small molecules as microarrays and detecting protein-ligand interactions en masse, J. Am. Chem. Soc. 121, 7967—7968 (1999). 

Kukar T, Eckenrode S, Gu Y, Lian W, Megginson M, She JX, Wu D. Protein microarrays to detect protein-protein interactions using red and green fluorescent proteins. Anal Biochem 2002 306 50-54. 

By using microarray technology, protein quantitative detection on one spot microarray is usually not possible. However, recently analysis of a microarray through laser ablation (LA)-inductively coupled plasma (ICP)-MS has been introduced, increasing the feasibility of protein detection (40). 

Electrical protein detection in cell lysates using high-density peptide-aptamer microarrays. J. Biol., 7,3. 

Kawahashi, Y., Doi, N., Takashima, H., Tsuda, C., Oishi, Y., Oyama, R., Yonezawa, M., Miyamoto-Sato, E., and Yanagawa, H. (2003) In vitro protein microarrays for detecting protein-protein interactions application of a new method for fluorescence labeling of proteins Proteomics 3, 1236-43. 

THE POTENTIAL OF PROTEIN-DETECTING MICROARRAYS FOR CLINICAL DIAGNOSTICS... 

We will focus on progress in detecting clinically useful diagnostic signatures and the development of protein-detecting microarrays to measure these signatures in the clinician s office. 

Fig. 2. A protein-detecting microarray. Each square in the grid represents a different feature of the array that would be impregnated with a particular protein ligand (blue shapes). When the sample is applied to the chip, each ligand will capture its target protein (orange and red coils in blow-up). The amount of target protein bound to each feature of the array would be quantitated with probes such as fluorescently labeled antibodies against the captured proteins. A fluoresence scanner would then measure the intensity of fluorescence (differently shaded green squares) at each spot, which would reflect the level of captured protein. (See Color Insert.)...

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