Lead Discovery Technologies

The first stage of target-based drug discovery is identification of molecules that affect the function of the target protein. This requires production of the pure target protein(s) and development of multiple assays to measure the binding of molecules to the target and the effect of those molecules on the target.

To obtain the proteins we can either use expression constructs from our large library of already-produced proteins, use constructs from collaborators, or use parallel efficient cloning strategies to create and test multiple new constructs for protein expression on a small scale, before developing methods for large scale production of the protein(s) in different forms suitable for different applications such as structural biology or assays. We are able to use E. coli, insect cell or mammalian cell expression systems, and have substantial capacity for protein purification using ÄKTA or other HPLC systems.

Most projects require several different assays to be developed or established in-house. We have done this for many different types of protein and we are well equipped for a large variety of biochemical assay techniques, with multiple liquid handlers and stacker-equipped plate readers giving capability for screening tens of thousands of compounds or hundreds of IC₅₀ measurements per day. For biophysical assays we have multiple different techniques available. We can also run assays on a high-throughput mass spectrometry platform. We have a selection of compound libraries available for screening.

Over the past 20 years these platforms have supported the determination of hundreds of novel crystal structures of human proteins as well as the discovery of dozens of high-quality chemical probes (potent and specific inhibitors that function in cells).

We can collaborate by providing a whole lead discovery cascade or through provision of individual services, proteins or assays. Our expertise and facilities are available through a small research facility (SRF). We can support you in assay development and miniaturisation or carry out protein production, assay development, screening campaign and lead characterisation for you.

Key people to contact

Dr. Jon Elkins (jon.elkins@cmd.ox.ac.uk, Academic Lead)

Dr. Ellie Williams (eleanor.williams@cmd.ox.ac.uk, SRF manager)

• Support with protein production for varied target classes
• Support with assay development for challenging targets, covering a wide range of technologies (see below)
• Assay miniaturisation to 384 well and 1536 well microtiter plate formats
• Conducting high throughput screening campaigns on our platforms
• Lead validation and characterisation with biophysical methods
• Time and training on our equipment if you have specific needs you can’t address in your own lab

Further information

Proteins of any type can be considered for production including integral membrane proteins, intrinsically disordered proteins and extracellular secreted proteins. Multiprotein complexes as well as LPS-free antigens and biomarkers can also be produced. Each year we have new collaborations within Oxford, with external academics and with industry.

Protein Production activities offered

• Construct Design
• Cloning and High-Throughput Cloning
• Test Expression (E. coli/Insect/Mammalian)
• Scale-up Expression (E. coli/Insect/Mammalian)
• Protein Purification

Protein expression and purification

• Labcyte Echo 525
• Shaker incubators for 96- or 24-well plates (Glascol)
• Shaker incubators for 50 mL to 3 L flasks (with CO2 for mammalian cells)
• ÄKTA pure, ÄKTA purifier and ÄKTA express FPLC instruments
• Dionex HPLC for analytical SEC / buffer screening
• LPS quantification

Compound storage and liquid handling

• Labcyte Echo 550 in Access workstation
• Formulatrix Tempest
• ThermoFisher Combi Mutidrop dispensers
• Roylan StoragePods

Multimode plate readers with plate stackers: BMG PherastarFS & FSX

• HTRF
• AlphaScreen/AlphaLisa
• Fluorescence intensity and polarisation
• Luminescence and absorption

Biophysical methods

• Surface Plasmon Resonance: BiacoreS200
• Biolayer Interferometry: FortéBio OctetRED384
• Microscale Thermophoresis: Nanotemper Monolith
• Differential Scanning Fluorimetry:
• Label free with NanoTemper Prometheus NT.48
• Dye based thermal shift assays with RT qPCR machines
• Isothermal Titration Calorimetry:
• TA Instruments NanoITC
• Malvern VP-ITC

Assays by mass spectrometry

• QTOF mass spectrometer (Agilent)
• Used with a high-throughput LC system (Agilent RapidFire) or a conventional HPLC (Agilent 1290 Infinity II)

Notable achievements

• Generation of high-throughput cloning and expression screening protocols enabling screening of >100,000 constructs in 16 years [1-6].
• Establishment of protocols and protein engineering to produce an array of epigenetic proteins including bromodomains and lysine demethylases [7-13].
• Development of expression systems and reproducible protocols to enable screening and production of integral membrane proteins [14-22].
• Creation of a toolkit of vectors for screening and production of proteins and strategies for identifying protein-protein interactions (Published shortly) [23].
• Production of a highly reproducible Spike antigen for use in a serology assay platform, developed at the University of Oxford, for identifying antibodies against SARS-CoV-2 [24, 25].