Aurelia Bioscience Ltd is a contract research organisation that provides services in biological assay development, biological screening and laboratory equipment consultancy to SMEs, universities, medical charities, pharmaceutical companies and screening equipment manufacturers as part of the early (pre-clinical) phase of the drug discovery process. Types of work will include fee-for-service bioassay development;fee-for-service compound screening; more specialised 'novel' assay development; medium to high-throughput compound screening (thousands to tens of thousands); screening cascade development i.e. multiple assays to add value to compound hits with, for example, a chemistry company; equipment consultancy with equipment manufacturers who wish to validate prototype equipment for commercial release.
Scientists at Aurelia Bioscience Ltd have over 100 years experience in drug discovery gained at several blue chip pharmaceutical companies including Hoffmann La Roche, Smith Kline Beecham, Glaxo-Wellcome, AstraZeneca, ICI and UCB.
They have worked in pre-clinical discovery, designing and developing low, medium and high throughput assays for hits-to-lead, lead identification and lead optimisation drug discovery phases in both cellular and non-cellular assay formats. They have experience of a wide range of technologies designed to prosecute these activities in a scalable manner whilst bringing to bear technological developments that have the ability to probe ever further into the molecular interactions from which the drugs of the future may be identified.
Research areas worked in include respiratory disease, inflammatory disease, oncology, CNS-pain, rare disorders and diabetes, working on a range of molecular targets in these areas.
Our scientists have in-depth experience of label-free technology applied to cell-based assays gained over the past 6 years using Corning Epic, PerkinElmer EnSpire and SRU Bioscience Bind instruments.
These systems incorporate an optical biosensor into a micro-titre plate to measure the cellular response to a stimulus. Specifically, the systems are able to detect changes in mass at the sensor surface resultant from alterations in cell shape and/or protein recruitment or migration to/from the membrane.
The technology is generic being independent of signaling pathway, it is non-invasive (no molecular biology required) and generates a kinetic response over seconds, minutes or hours resulting in an information rich readout.
Label-free techniques are highly suitable for measuring responses in native or physiologically relevant systems such as primary human and animal cells without the need for over-expression or amplification through genetic modifications. This is of great benefit in understanding the molecular pharmacology of a receptor in its native environment, whereas a recombinant system does not necessarily have the natural coupling or utilise the correct signaling pathway.
Label-free techniques may also be of benefit where there is no simple/standard biological readout e.g. for de-orphanisation of receptors and identification of pharmacological “tool” compounds.
We have used FLIPR in a range of assay formats, readouts and target types including:
Antagonist and agonist screening in recombinant cell based assays
Working within 7-transmembrane receptor (7-TMR) receptor (also known as G-Protein Coupled Receptors or GPCR) biology for the last 15 years we have detailed knowledge and experience of developing assays and screening for 7-TMR activation and inhibition. We have used fluorescent methodologies with Fluo-3, Fluo-4, Fluo-4 no wash reagents and Fluo-5 in 96, 384 and 1536 well plates.
The 8200 Cellular Detection System (Fluorescent Microvolume Assay Technology – FMAT) from Applied Biosystems is a laser scanning macro-confocal fluorescent plate reader.
FMAT scans the central 1mm2 area of a micro well to a depth of 100μm from the base of the well using a red 633nm Helium-Neon laser to identify “objects” within the focal plane. Objects such as cells or beads and associated fluorescence are detected and measured by two photomultiplier tubes (PMT1, 650-685 nm; PMT2, 685-720 nm) simultaneously. Fluorescence not associated with objects is undetected, so that the signal-to-noise ratio is dramatically increased. In addition, a 633nm excitation source reduces interference from the auto-fluorescence of compounds and plastic.
These features enable FMAT to perform homogeneous assays without the need to remove unbound label using time consuming washing steps that introduce significant assay variability.
Cell-based assays may be conducted on live or fixed cells with both adherent and suspension cell lines. Bead-based assays are also easy to perform and existing ELISAs are readily converted to homogeneous FMAT formats (FLISA) by coating beads with capture antibodies (Ref – J Biomol Screen March 2008 vol. 13 no. 3 210-217).
High Content Screening (HCS) uses microscopic imaging to observe, measure and analyse multiple parameters within cells in a biological assay.
By using fluorescent dyes, fluorescently labelled proteins and fluorescently labelled antibodies, events that occur within cells following the addition of compounds and ligands can be detected and quantitated.
In addition to our in-house CellInsight CX5 platform, our scientists have access to the Image Xpress Ultra from the Molecular Devices Corporation.
Aurelia Bioscience have invested in a Wes from ProteinSimple so as to provide a reliable automated western blotting service to clients.
Simple WesternTM Assay: The gel-free, blot-free, hands-free reinvention of Western blotting
Whilst Western blotting remains the pre-eminent method of choice for protein detection and characterisation, the various improvements to individual aspects of Western work flow developed in recent years have had little impact on the relatively poor reproducibility, lack of accurate quantification, extensive time to result and reliability issues associated with traditional Western blotting.
The WesTM system from ProteinSimple is a fully automated device which runs the Simple Western size assay; a highly reproducible and sensitive alternative to standard Western blotting assays. Wes employs capillary electrophoresis combined with chemi-luminescence based antibody detection to run the entire Western immunoassay process from start to finish, providing dramatic improvements in sensitivity, reproducibility, quantitative accuracy and time to result compared to all other legacy Western blotting methods. Simple Western assays cover three separation ranges: 2 – 40kDa, 12 – 230 kDa and 66 – 440kDa.
Wes delivers exceptional results for virtually all sample types requiring a Western blotting approach. Minimal sample volume (3-5uL) at a low total protein concentration (0.2ug/uL) are required. Runs typically comprise 24 samples plus a size ladder and full results are generated in just over 3 hours. The Simple Western assay uses quality controlled proprietary consumables and all the client would ordinarily need to supply are their samples and a suitable primary antibody. An ever expanding wide range of Simple Western certified primary antibodies are available and primary antibodies from other sources may also be used.
In addition to Simple Western Immunoassay, Wes also runs the Simple Western Total Protein assay
Cell based assays are of key importance at a number of stages in the drug discovery process.
Being able to examine the interaction between your target and compounds in a cellular context is of great advantage in:
The team at Aurelia Bioscience have developed and screened using cell-based assays for over 15 years. Our knowledge and skills in cell culture and manipulation, combined with our expertise in developing cell-based assays allow us to work rapidly and efficiently on the prosecution of your target in appropriate assays.
Our core technology platform enables us to measure readouts such as fluorescence, luminescence and chemi-luminescence.
Example assays include but are not limited to:
With over 20 years of screening experience we are adept at designing screening assays and screening 100,000 to 1 million compounds in 384 well and 1536 well plates.
We have screened up to one million compounds in both cell and biochemical assays using a number of read outs including FLIPR, FMAT, FI, FRET, chemiluminescence, luminescence and label-free formats.
We have extensive experience of cell-based assays in HTS where we have used either cell lines expressing the target in a recombinant manner, Bacmam transient expression and cryopreserved cells.
For human primary cell screening we have used either cryopreserved or freshly isolated cells such as PBMC’s and neutrophils. We understand the importance of delivering robust, reproducible assays with excellent quality control statistics for use in high throughput screening.
We are able to use our experience to assist your drug discovery projects in a number of ways:
Screening densities will be utilised which are consistent with the assay format and the number of compounds to be screened.
Our team has extensive experience of developing and screening compounds in biochemical assays for a range of targets including but not limited to proteases, kinases, and phosphodiesterase enzymes and protein:protein interaction.
Our core technology platform enables us to offer fluorescence (Fluorescent Resonance Energy Transfer [FRET], Fluorescent Intensity [FI], Time-Resolved Fluorescence [TRF]), AlphaScreen based chemiluminescence, luminescence and absorbance read outs to develop the best assay for your target needs.
Throughput is based on 96, 384 or 1536 well format giving the client the option to screen from 10’s to 100’s to 1000’s to millions of compounds. Screening approaches can be replicates of a single concentration for hit identification or multiple concentrations to generate assay response curves for structure activity profiling and compound progression.
We offer flexible services developing new assays or adapting existing assay to meet your needs e.g. incubation time, kinetic versus endpoint measurements etc.
Aurelia Bioscience is now able to offer a qPCR service to our clients. Using our Applied Biosystems™ QuantStudio™ 6 Flex Real-Time PCR system, we are able to investigate quantitative differences in mRNA expression in relation to pathways and targets identified as being of interest to clients. The QuantStudio™ offers a high throughput real-time PCR platform (with a 384 well block) with sophisticated data analysis software. Designed to easily manage large data sets, the embedded gene expression study application can analyse >100 runs, while heat maps and scatter plots provide quick checks on data quality.
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