Etude de l'interaction médicament/récepteur par spectrométrie de masse : mise en place et validation de nouveaux protocoles de criblage moléculaire

Abstract : Discovering new drugs by biomolecular screening is a central task of pharmaceutical research. Mass spectrometry, as a reliable, reproducible, sensitive and specific technique, compatible with a wide range of samples and offering an excellent throughput, shows its potential in different strategies of research and development. The aim of this work was to develop and validate a new strategy, involving matrix assisted laser desorption/ionization coupled to time of flight mass spectrometry (MALDI-TOFMS) to screen the ability of different compounds, including plant extracts, to bind to two biological targets (tubulin and DHFR). The protocol is therefore divided into three main steps : an incubation of the compounds to be tested with target, an elimination of all unbound compounds and the MALDI-TOFMS detection of target-bound compounds. Our protocol was validated and the results that can be obtained were discussed. The throughput offered by this technique was evaluated as 60 samples in 1h50 to 3h30, or 18 to 32 samples per hour. Finally, we developed a new approach to perform a secondary screening of active compounds. The protocol was applied to screen crude plants extracts (colchicum autumnale, catharanthus roseus and green tea) and allowed to find 20 tubulin-binding or DHFR-binding molecules, and the relative affinity of one of these was also evaluated
Document type :
Theses
Complete list of metadatas

Cited literature [15 references]  Display  Hide  Download

https://hal.univ-lorraine.fr/tel-01752607
Contributor : Thèses Ul <>
Submitted on : Thursday, March 29, 2018 - 1:52:22 PM
Last modification on : Monday, April 16, 2018 - 10:42:24 AM

File

Hannewald_Paul_SMZ0842.pdf
Files produced by the author(s)

Identifiers

  • HAL Id : tel-01752607, version 1

Collections

Citation

Paul Hannewald. Etude de l'interaction médicament/récepteur par spectrométrie de masse : mise en place et validation de nouveaux protocoles de criblage moléculaire. Autre. Université Paul Verlaine - Metz, 2008. Français. ⟨NNT : 2008METZ042S⟩. ⟨tel-01752607⟩

Share

Metrics

Record views

21

Files downloads

31