A set of chemical biology resources were built by the Nodwell Lab for the detection and purification of biologically active small molecules from various biological sources. In principle this infrastructure can be used to isolate any compound for which a suitable assay is available. The laboratory is a BSL2 facility so sources can include pathogenic bacteria, fungi, and cancer cells.
The typical work flow in the lab involves the chemical extraction of cells using any of several volatile solvents. The extract is concentrated by evaporation using a rotavap (Heidolph Hei-vap) or Genevac (EZ 2.3-Elite) system. Aqueous extracts are dried using a lyophilizer (Labconco Freezone). The resulting extracts are partitioned into individual components by liquid chromatography.
Instruments
LC/MS – Waters Acquity UPLC/Xevo G2-S QTof
Liquid chromatography – Mass spectrometry
The device allows the separation of small molecules by reversed phase chromatography and the detection of peaks by MS, MS-MS, and by broad-spectrum light absorption. It is typically used for analytical scale investigation of small molecules.
HPLC – Waters Alliance
HPLC
This allows the analysis and purification of compounds on an analytical and semi-preparative scale with the use of a photodiode array detector and automated fraction collector.
Flash Chromatography – Reveleris X2
Flash chromatography
This instrument can collect fractions on a preparative scale using both integrated UV detector and ELSD (for non-UV absorbing molecules) and can switch between normal phase and reversed phase chromatography.
Training
All of this apparatus requires extensive training to be used independently, including BSL2 certification.
The resources can also be accessed through collaboration with members of the Nodwell Lab.
Location
MaRS Center, West Tower
661 University Ave., 16th Floor
Contact
Publications
Arryn Craney, Cory Ozimok, Sheila Pimentel-Elardo, Federico Capretta and Justin R. Nodwell (2012). Chemical Perturbation of secondary metabolism demonstrates important links to primary metabolism. Chemistry and Biology 19: 1020-7
Leslie Cuthbertson, Sang-Kyun Ahn and Justin R. Nodwell. (2013). A mechanism of inducible antibiotic resistance revealed using a global relational tree for one-component regulatory systems. Chemistry and Biology 20(1): 20: 232-40
Salman Ahmed, Arryn Craney, Sheila Pimentel-Elardo and Justin R. Nodwell. (2013). A synthetic, species-specific activator of secondary metabolism and sporulation in Streptomyces coelicolor. Chembiochem 14(1): 83-91.