Skip to Main content Skip to Navigation
Theses

IMAGERIE PAR RESONANCE DES PLASMONS DE SURFACE POUR L'ANALYSE SIMULTANEE DE MULTIPLES INTERACTIONS BIOMOLECULAIRES EN TEMPS REEL

Emmanuel Maillart 1
1 Laboratoire Charles Fabry de l'Institut d'Optique / Macsybio - bio
LCFIO - Laboratoire Charles Fabry de l'Institut d'Optique
Abstract : The current growing requirements, in the field of biomolecular interactions analysis, require the development of automated high throughput systems like fluorescence microscopy that remains the method of choice. Although direct methods (without labeling of the molecules to be analyzed) have a lower sensitivity than those using labels, their many advantages (real time monitoring, simplification of the sample preparation) are of importance for biomolecular interactions quantitiative analysis. Moreover, the surface plasmon resonance imaging allows simultaneous measurements of many interactions like the methods with labels. To implement such a device, we identified its various critical parameters, likely to be improved, like the nature of the metal layer. The use of the thus developed sensor relies largely on surface chemistry: we have chosen the electrochemical synthesis of polypyrrole films that allows, in a controlled and structured manner, a simple and robust functionalization of spots containing various molecules. In parallel, an adequate home-developed software, including acquisition of images whose processing allows the real time monitoring of the biomolecular reactions occurring on each spot and the extraction of their association rate, dissociation rate and affinity constant between molecules. Lastly, we demonstrated the application possibilities of this system to the stufy of hereditary diseases or cancer through DNA/DNA (K-ras), DNA/protein (p53), protein/protein (hCG) and oligosaccharide/protein interactions (HP6/SDF-1).
Complete list of metadatas

Cited literature [64 references]  Display  Hide  Download

https://pastel.archives-ouvertes.fr/tel-00006480
Contributor : Emmanuel Maillart <>
Submitted on : Friday, July 16, 2004 - 3:36:30 PM
Last modification on : Wednesday, October 14, 2020 - 4:00:35 AM

Identifiers

  • HAL Id : tel-00006480, version 1

Citation

Emmanuel Maillart. IMAGERIE PAR RESONANCE DES PLASMONS DE SURFACE POUR L'ANALYSE SIMULTANEE DE MULTIPLES INTERACTIONS BIOMOLECULAIRES EN TEMPS REEL. Biophysique [physics.bio-ph]. Université Paris Sud - Paris XI, 2004. Français. ⟨tel-00006480⟩

Share

Metrics

Record views

1433

Files downloads

4993