Purdue University houses world-class facilities to support life-science research. The Flaherty lab is located in the Robert Heine Pharmacy Building and his research utilizes facilities within the Purdue Institute for Drug Discovery. The lab has equipment to support organic synthesis and purification, protein expression and purification, and binding experiments. The lab also benefits from campus cores such as the Purdue Interdepartmental NMR Facility, Campus-wide Mass Spectrometry, Biophysical Analysis Laboratory and the Purdue Chemical Genomic Facility. Lab members have access to and utilize these resources on a daily basis.
Laboratory Space in RHPH
The Flaherty lab is located in room 403 in RHPH. The lab is equipped with seven fume hoods and bench space for organic synthesis. Additional bench space is dedicated to protein expression and purification to support our research projects. An instrumentation room adjacent to the lab houses an rt-PCR instrument for performing protein thermal shift binding experiments and an HPLC for small molecule purity analysis and semi-prep purification. A student office is located next to the lab with computers for molecular modeling/computational docking experiments and common space for lab discussions.
Flaherty Lab Equipment
Yamazen W-Prep 2XY Flash Chromatography System
This flash chromatography purification system allows has two channels that allows for simultaneous purification of organic small molecules in parallel or in tandem. One channel is routinely operated in reverse-phase mode while the other in normal-phase mode. It has a dual channel fraction collector that identifies peaks and collects fractions automatically without the chemist present. This system cuts down significantly on the amount of time it takes to isolate a pure small molecules.
Biotage Initiator Microwave Synthesizer with Robot 60
This instrument is used for microwave synthesis of small molecules. It can be programed for automated synthesis of several samples in succession using the robot 60. We also have the peptide work station that cuts down on the time between steps for solid-phase peptide chemistry.
Agilent 1200 Series HPLC
This HPLC system is used for purity analysis of small molecules synthesized in the lab. It has an autosampler for set up of multiple samples in succession. The system is also used for semi-prep HPLC purification if necessary.
Protein Expression and Purification Station
Our lab has a need for recombinant protein to support biochemical/biophysical binding assays, crystallography, and NMR binding experiments. This station has everything necessary to recombinantly express protein in E. coli, isolate, and purify for use in our experiments, including and incubator/shaker, centrifuge, peristaltic pump, and SDS-PAGE gel systems.
AKTA PURE FPLC Station
This FPLC allows for size-exclusion chromatography purification of proteins to produce ultra-pure samples for all in vitro experiments, crystallography and NMR. We have two size-exclusion columns to isolate proteins of various sizes.
This system also allows for preparatory reverse phase chromatography purification of peptides and small molecules.
Applied Biosystems StepOne Plus PCR
This system is utilized to perform protein thermal shift experiments on our targets. We use this assay to identify small molecule binders and to measure the stability of proteins that have undergone mutagenesis. The station has the Protein Thermal Shift software license from Applies Biosystems to provide an intuitive analysis package for the experiments that are performed.
*Are lab assists those interested int using protein thermal shift for screening of small molecule binders.
Small Molecule Libraries
The lab maintains two in-house small molecule libraries. First, a 30,000 small molecule library that was selected from the DIVERSet molecules from ChemBridge. Second, a 1,500 molecule fragment library selected from the Life Chemicals Fragment Collection.
*Both libraries are available for projects with selected collaborators. Contact Dr. Flaherty for more information.
Computational Modeling Computers
Our lab has two computers dedicated to compound library curation, cheminformatics and computational docking using the Schrödinger Small Molecule Drug Discovery Suite. Both have additional processing power to handle large, computationally demanding jobs.
Departmental and Campus Equipment
Advion Compact Mass Spectrometer with TLC Express
and ASAP probe
Our lab is a major user of the Advion CMS for small molecule, peptide, and macromolecule analysis. The mass spec has both an ESI and APCI source that may are interchangeable in a matter of minutes. It is equipped for three types of sample injections. First, the TLC Express front end allows users to perform a standard TLC for reaction monitoring and then place the TLC plate in the TLC Express. This then lowers a head onto the TLC plate
NanoTemper Microscale Thermophoresis
Our lab has become a major user of the MST instruments available in the Purdue Center of Chemical Genomics. The instrument monitors macromolecular movement in solution across a thermal gradient. This can then be used to assess changes in thermophoresis in dose-response to obtain binding affinity measurements in as little as 15 min with 20 microliter of sample.
ForteBio Octet Red 384
The Octet Red, housed in the Biophysical Analysis Core Lab in Bindley BioSciences Center, allows for binding kinetic measurements in a well plate format. Sensors are loaded with a protein and then dipped into wells containing ligands or macromolecules to measure binding affinity and kinetics.
Purdue Interdepartmental NMR Facility
Our lab frequently uses a Bruker ARK300 and ARX500 housed in RHPH as well as a Broker Avance III 800 with cryoprobe housed in Brown Chemistry Building.