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Label-free microarrays for the parallel analysis of small (bio)molecules


Microarrays- flat surfaces that bear hundreds or thousands compounds, each of which able to recognize a complementary molecule- are mostly used today in nucleic acid analysis, but the technology is not developed in the screening of small drug-like (bio)molecules.

We propose to exploit the combinatorial parallel approach for the fabrication of small (bio)organic molecule microarrays on glass support, and to adapt the technique in order to screen directly the molecules for biochemical activity. The main innovation of the project is the original idea to attach both small (bio)molecules and fluorophores onto a glass substrate in the fabrication of the microarrays.

The fluorophore, which plays the role of an internal read-out system, will allow a direct screening b y fluorescence microscopy without extra labels. The arrays will be prepared using either light-directed or microcontact imprinted fabrication. Using the first method, glass substrate functionalized with photodegradable protecting groups will be modified on specific areas by using photolithography masks.

The areas deprotected with light can be reacted with first a (bio)molecule, secondly a fluorophore on the same site. Next, light is directed to another region of the substrate using a second mask, and the chemical cycle is repeated with a different molecule and a different fluorophore. The second method involves the use of a polymeric stamp coated with a solution of (bio)molecule or fluorophore, which is pressed onto the functionalized glass substrate.

Molecules are transferred only at the place of direct contact between the stamp surfaces and the substrate. Screening of these microarrays upon addition of an analyte can then be performed simply using fluorescence microscopy. This new approach allows faster and cheaper fabrication of a new class of microarrays on glass. It does not only increase the speed of the analysis but also allows recycling of the microarrays for different targets.

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University Of Twente