COMBIARRAY · Label-free microarrays for the parallel analysis of small (bio)molecules
FP6 — Marie Curie Actions (Human Resources and Mobility)
- Duration
- 2006-01-01 → 2007-11-30
- EU contribution
- €148,710
- Participants
- 1
- Scheme
- EIF
Lines connect the coordinator with its partners. CORDIS does not always give exact coordinates for projects before 2014. These points are placed at city or country level.
Results in brief
Final Activity Report Summary - COMBIARRAY (Label-free microarrays for the parallel analysis of small (bio)molecules)
Combiarray aimed at designing and testing new fluorescent sensors for analysing specific biological interactions. The project has been concentrated mainly on the detection of DNA hybridisation, because of its potential applications in DNA microarrays. Nowadays, fluorescent DNA microarrays need a labelling step of the target molecule, which we would like to avoid by creating label-free fluorescent sensors. In addition, in the course to reduce the fabrication's costs of a sensor, we focused on the use of microcontact printing, a cheap soft lithography technique, for the deposition of DNA strands. Synthetic oligonucleotides have been covalently attached to reactive self-assembled monolayers on glass surfaces, using microcontact printing deposition. The fabrication conditions have been studied and optimised in order to take into account i) the resistance of the surface to aqueous media, ii) the printing of polar molecules like DNA using a very apolar material. Surfaces functionalised with oligonucleotides have been hybridised with complementary or non-complementary strands, and the process has been followed using labeled oligos and fluorescence microscopy. The label-free sensing of hybridisation has been realised by using a fluorescent intercalator as the sensor molecule. For this purpose, non-labeled oligos have been deposited to glass surfaces, hybridised with complementary strands, and subsequently interacted with ethidium bromide to evidence the presence of single or double strands of DNA.
Data: CORDIS, © European Union
Project objective
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.
Original text from CORDIS.
Participants
- UNIVERSITY OF TWENTE SUPRAMOLECULAR CHEMISTRY AND TECHNOLOGY · ENSCHEDECoordinatorCity levelNetherlands
Links
Data: CORDIS, © European Union