Publication and poster page

3rd Cell-Based Assay UK Conference (organised by VisionGain) - oral presentation...

"The Role of Phenotypic and Orthogonal Screening in Early Drug Discovery"

For the full article click here

For other interesting cell-based assay presentations from the conference click here

Society for Laboratory Automation and Screening (SLAS - 2013) - oral presentation...

"Orthogonal and Phenotypic Approaches to Drug Discovery"

The presentation covers experimental work looking at multiplexing two assays in one well using the same cells. Label-free kinetic measurements from cells treated with agonists and/or antagonists are combined with beta-arrestin measurements and calcium measurements from the same cells.

 For the full article click here.

Society for Laboratory Automation and Screening (SLAS - 2013) - poster...

"Phenotypic screening meets target-based drug discovery: Multiplexing label-free cellular measurements with DiscoveRx assays on the same cells to aide mode of action studies – two bangs for one buck!"

A recent publication (Swinney, D & Anthony, J., Nature Reviews: Drug Discovery, vol 10, July 2011 p 507) reviewed historical data and concluded that phenotypic approaches can be more successful than target-based discovery. The use of label-free optical biosensors for studying integrated cellular responses as a phenotypic approach to drug discovery has increased over recent years. A key benefit is the high sensitivity in endogenous systems and primary cells. In an attempt to bridge the gap between phenotypic and target-based screening we have used the DiscoveRx PathHunter CHO-K1 Histamine H1 receptor beta-arrestin cell line to examine the feasibility of multiplexing a cell-based label-free readout with other second messenger and downstream signalling assays e.g. (beta-arrestin) and calcium flux. A significant advantage of label-free measurements is that it is non-invasive, allowing the same cells to be used in a second assay format. This allows mode of action, deconvolution and toxicity studies to be performed on the same cells that generated the label- free responses. Here, we describe the steps taken to identify suitable assay conditions that enabled both label-free and secondary assays to be conducted on the same cells in the same wells of the assay plate performed using the EnSpire multimodal reader from PerkinElmer.

 For the full article click here.

Paper: Journal of Biomolecular Screening article (2009) on Label-free technology

"A 100K well screen for a muscarinic receptor using the Epic label-free system--a reflection on the benefits of the label-free approach to screening seven-transmembrane receptors”.

Seven-transmembrane receptors (7TMRs) are a family of proteins of great interest as therapeutic targets because of their abundance on the cell surface, diverse effects in modulating cell behavior and success as a key class of drugs. We have evaluated the Epic label-free system for the purpose of identifying antagonists of the muscarinic M3 receptor. We compared the data generated from the label-free technology with data for the same compounds in a calcium flux assay. We have shown that this technology can be used for high throughput screening (HTS) of 7TMRs and as an orthogonal approach to enable rapid evaluation of HTS outputs. A number of compounds have been identified which were not found in a functional HTS measuring the output from a single pathway, which may offer new approaches to inhibiting responses through this receptor

For full article click here

Oral presentation ELRIG (2007)

"General overview and application of label-free technology in screening"

For full article click here

Society of Biomolecular Screening Conference (2006)

Poster: "AstraZeneca Evaluation of EPIC Label Free GPCRs"

For full article click here

Poster: "Development of a 384-Well Small Molecule Binding Assay for Trypsin on the Epic Label-Free Reader"

For full article click here

Article: "Current Trands in Label Free Technology"

The implementation of automated systems, increased sensitivity and the development of new technologies have recently broadened the use of label-free methods from primary screening through to mechanism-of-action studies in lead optimisation. The trend is now towards adopting label-free methods earlier in the drug discovery process so that compounds can be tested against native protein or cells resulting in improved biological assessment of compounds more predictive of therapeutic effect.

Healthcare reform, tougher regulatory hurdles and patent expiry of drugs are putting the pharmaceutical industry under increasing pressure, with R&D costs continuing to increase but numbers of NMEs remaining static. It is clear that for companies to remain competitive, they not only need to reduce project attrition and improve cycle times but also identify technologies that provide more physiological mechanistic data to help identify compounds that translate in vitro activity to................

For full article click here

Poster: “Solubility Rules OK. What Ultrasonic mixing can do for you”

Poster using acoustic mixing technology developed by Microsonic systems used to solubilise compounds and increase the degree of mixing in small volume assays.

For full article click here

Paper: Journal of Biomolecular Screening:

“Identification of iNOS Inhibitors Using InteraX™” 

Inducible nitric oxide synthase (iNOS) is active as a homodimer. A cell-based assay suitable for high-throughput screening (HTS) was generated to identify inhibitors of iNOS dimerization using the InteraX™ enzyme complementation technology of Applied Biosystems. The cells contain 2 chimeric proteins of complementing deletion mutants of β-galactosidase, each fused to the oxygenase domain of human iNOS. The assay was characterized using known iNOS dimerization inhibitors, which gave a decrease in β-galactosidase activity. Surprisingly, the assay was also able to identify compounds that have the same profile as known inhibitors of fully formed dimeric iNOS by causing an increase in β-galactosidase activity. The iNOS InteraX™ assay was used to screen ~800,000 compounds in a 384-well format. After hit confirmation, 3359 compounds were taken forward for full IC₅₀ determination in InteraX™ and cytotoxicity assays. Of these compounds 40.5% were confirmed as greater than 10-fold more active in InteraX™ compared to a cytotoxicity assay and were classified as potential iNOS dimerization inhibitors as they did not inhibit β-galactosidase alone. In the same primary screen, 901 compounds gave a significant increase in β-galactosidase activity. Many of these were known inhibitors of iNOS. After IC₅₀ determination in InteraX™ and cytotoxicity assays, 182 novel compounds remained as potential arginine-competitive inhibitors of dimeric iNOS. (J Biomol Screen March 2009 vol. 14 (3), 263-272)

For full article click here

Webinar for Labcyte Inc

"The use of primary cells in HTS"

Presentation on the use of acoustic dispensing into 1536 well plates for primary human neutrophils screening in a GPCR calcium mobilisation assay.

For article click here

Poster: "AlphaScreen assay for high throughput screening of a protein:protein interaction"

For article click here

Article: "The use and impact of automated cell culture systems to supply cells for high throughput screens"

For article click here:

Oral Presentation: 6th Molecular Devices user group meeting on 1536 FLIPR screening

For article click here:

Oral Presentation: The Use of Human Primary Cells in HTS: A Reality in the Making Using FLIPR^TETRA in 1536 Well Format

For article click here