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High-Content Screening Surges Ahead

Last Updated on Tuesday, 1 December 2009 11:33 Written by Editor Tuesday, 1 December 2009 11:33

High-content screening (HCS) and the technology to do it faster, on more compounds in a shorter period of time, and to generate quantitative, multiparametric data took center stage at CHI’s “High Content East” meeting held in Boston last month. Presenters described how they are implementing enhanced screening systems, image-analysis methods, and data-management strategies to achieve daily HCS runs on tens of thousands of wells and screening campaigns totaling 200,000 to 3 million wells.

High throughput HCS—albeit not yet reaching the numbers common for conventional high-throughput screening (HTS) and with lingering limitations and challenges related to live-cell imaging over time—is making its mark and being used to probe the biological basis of disease and to detect even subtle phenotypic changes in response to experimental compounds.

Determining whether a cell looks like a cancer cell, for example, typically requires being able to detect subtle morphological changes, such as small alterations in size or structure, changes in the connections a cell makes with neighboring cells, or variations in the texture of staining. These have, historically, been mainly qualitative parameters detected by studying and comparing images of cells.

In her talk at the conference, Anne Carpenter, Ph.D., director of the imaging platform at the Broad Institute of Harvard University and MIT, presented her group’s work using HCS and image analysis to quantify difficult phenotypes and differentiate disease states such as leukemia.

Not only do HCS systems and image-analysis software automate the screening process, enabling theanalysis of many more cells in less time and increasing the chances of detecting even small numbers of altered cells, they can also utilize algorithms that evaluate defined combinations of parameters in a quantifiable manner and apply techniques to distinguish between clumping or closely juxtaposed cells. Relying on computer-based image analysis also standardizes the process, eliminating factors such as variability in human expertise and experience, consistency, and fatigue.

Dr. Carpenter’s group uses machine-learning methods to train image-analysis software to identify subtle phenotypic changes. Biologists work with the software in an iterative fashion in a process called supervised machine learning. They teach and correct the computers on a series of test images, refining the system’s knowledge base in a process that typically takes less than a day. The group developed the algorithms used by the biologists and has made them available as open-source software.

A recent paper published in PNAS by T. R. Jones, et al., documents the use of a trained image-analysis system to discriminate 15 different cellular phenotypes. Other projects involve teaching the software to discriminate leukemic from normal cells, to identify liver cells that are growing normally in culture—to aid in the development of physiologic models of liver function for use in drug testing—and training computers to detect subtle changes that signal the initiation of cell division for studying cell-cycle regulation in cancer.

Neil Carragher, Ph.D., senior scientist in the advanced science and technology laboratory at AstraZeneca, described how the company is applying high-content and live-cell imaging techniques and integrating the results with data derived from in vivo imaging and proteomic studies to improve clinical predictability.

Dr. Carragher’s group combines the results of high-content in vitro and in vivo assays to generate mechanistic information about phenotypic responses on candidate therapeutic compounds. The goal is to create a multiparametric fingerprint of a phenotype from images generated by HCS and to use this knowledge to enhance predictions of efficacy and toxicity early in drug discovery and reduce attrition later in development.

The phenotypic signatures are based on measurements of approximately 150 different parameters per cell for each assay. Data from multiple assays is collated for every test compound and compared with data obtained using well-characterized reference compounds to generate mechanistic hypotheses.

Only recently has open-source and commercial software become available “that allows you to quantitate more complex phenotypes, subtle changes, and heterogeneous responses from images,” Dr. Carragher said.

His group is employing two main approaches—each with different advantages and limitations. The first strategy relies on Definiens’ Cognition Network Technology™ software that allows users to develop algorithms that capture, computationally, what researchers can see visually. “It is very much context-based” and identifies objects based on how they are related to others in the image, rather than as individual pixels, explained Dr. Carragher. The in-house algorithm-development process depends on iterative programming steps. The other approach involves machine-learning tools using software such as the CellProfiler developed at the Broad Institute.

Redirecting Approved Drugs
Identifying new applications for FDA-approved drugs using HCS and image-based systems biology is the focus of work being done by Stephen Wong, Ph.D., founding director of the bioinformatics and biomedical engineering program and the cellular and tissue microscopy core at the Methodist Hospital Research Institute and professor of radiology and neurosciences at Weill Cornell Medical College.

Dr. Wong gave examples of screening campaigns to decipher targets in the pathways responsible for the metastasis of breast cancer to the brain in his talk. He specifically described the computational tools his group is developing for high-content and network analysis, and the animal-imaging techniques being used to evaluate combinations of small molecule chemotherapeutic agents for their ability to cross the blood-brain barrier and to have an effect against central nervous system metastases in breast cancer.

Dr. Wong’s group has also developed a series of quantitative image-analysis tools, including zebrafish image quantifier (ZFIQ), as well as software for studying neuronal spines (NeuronIQ), neurites (Neurite IQ), and time-lapse mitotic events in cells (DCellIQ). Dr. Wong’s HCS/systems biology research is funded by the NCI, NIA, and NLM.

Because the compounds being studied are already approved drugs, Phase I trials are not needed. The quantitative data generated from HCS provides the evidence necessary for moving into Phase II studies, shortening the drug-development cycle to a year or less.

The types of studies essential to Dr. Wong’s efforts, such as assays to monitor cell-cycle regulation or dendritic spine dynamics, require time-lapse, live-cell imaging. Looking at fixed cells provides only an artificial snapshot of where cells are at a particular point in time, explained Dr.  Wong. “We want to look at a 384-well plate of continuously growing cells over five to six days,” he said, and in his view none of the instrument manufacturers competing in the HCS market has yet to provide a robust, incubator-based, environmentally controlled system that can achieve this.

Vendors have tended to view HCS as just another type of high-throughput screening, but live-cell imaging done in as natural an environment as possible has quite different requirements, contended Dr. Wong.

“Vendors are going in the wrong direction. The power of HCS is in the ability to visualize things in action and to extract lots more quantitative information from the images. If you, instead, retrofit HCS to HTS, you are losing its advantages,” such as the ability to see cells or spines change over time, to visualize cell-cell interactions, and to sync cell populations and study cell-cycle events in time-lapse, said Dr. Wong.

In any experiment, “if you generate enough data you will get hits, but how many will be real hits versus false positives?” asked Dr. Wong. “We need to push the quality upfront on the biology side” and screen out, earlier in the discovery process, compounds that are destined to fail.

Researchers at Pfizer are using HCS to study the genetic variation and physiologic interactions that underlie hepatic insulin resistance in type 2 diabetes and the prediabetic state. Diabetes is a complex, multigenic disease, and while advances in genomic and SNP-based technologies have led to the identification of at least 30 genes that contribute to the diabetic phenotype, much work remains to understand their role in cell biology and disease and how they interact.

“If you are careful about the cell models you choose, you can use HCS to characterize these genes and monitor their effects on biochemical pathways,” said Steven Haney, Ph.D., associate fellow in biological profiling at Pfizer’s biotherapeutics and bioinnovation center. The company has invested heavily in developing cell models that are representative of human physiology, including hepatocytes that faithfully mimic liver function when grown in culture.

The other main aspect of this research effort involves identifying changes that affect the diabetic phenotype, specifically glucose storage and utilization pathways,  and distinguishing between effects that involve the insulin-signaling pathway from more general phenomena related to activation of toxicologic or stress pathways.

“HCS can alert us to things we don’t necessarily know to look for, in a mechanism-independent way,” said Dr. Haney. “The increasing throughput of HCS allows us to look at a lot of cells and determine whether subtle phenotypic changes are significant or spurious.”

Vendors Roll Out Image-Analysis Solutions

Versatility across application areas, from microscope-based imaging for detecting intracellular phenomena to high-speed scans at the cellular level to whole organism screening, is the focal point of instrument development at MDS Analytical Technologies. “With the options in our Complete Solution and the right infrastructure, you can use image-based assays for primary screening. We have tackled all the common bottlenecks,” said Michael Sjaastad, Ph.D., director of marketing for cellular imaging at MDS.

The IsoCyte® DL laser-scanning cytometer complements the company’s ImageXpress® instrument platform as part of its overall HCS solution. MDS offers a high-throughput option that can screen and do image analysis on a 1,536-well plate in two to five minutes, according to Dr. Sjaastad. The instrument can image whole wells for accurate cell counting in cell-viability measurements, scan a microscope slide, or produce and analyze images of organisms such as zebrafish or worms when used in conjunction with the MetaXpress image-analysis software.

For now, current systems “have the image resolution and acquisition speed researchers need,” and in Dr. Sjaastad’s view, future improvements will focus on “streamlining the data-analysis workflow and bringing the costs down per data point.”

In a workshop at the meeting, Oliver Leven, Ph.D., head of screener professional services at Genedata, identified several ongoing challenges in HCS, including managing the volume and complexity of the data, improving the efficiency of data analysis, and creating an audit trail of results interpretation. As the throughput and scale of HCS increases, so too, do the difficulty and scope of these challenges.

As researchers scale up an assay for high-throughput HCS, they need to select a defined set of parameters that represent the phenotype of interest and that allow them to assess the quality of both the assay and the data output. They also need to identify threshold values above or below which a result signifies a change in phenotype.

The typical HCS image-analysis software that drives HCS systems routinely quantifies the cell images to yield a numerical description of the phenotypes. For large experiments, however, Dr. Leven described the researcher’s need to go back and view an image associated with an interesting or suspicious measurement as a persistent bottleneck.

“The image is the experiment,” said Dr. Leven. A hit should signify a change in the cells, but it could also be an anecdotal finding or the result of an image out of focus. Distinguishing true hits from false positive results remains a challenge.

Dr. Leven recounted the HCS projects  that Genedata has performed for its pharma customers emphasizing the ability of the company’s High Content Analyzer—a new addition to the Genedata Screener® enterprise solution—to retrieve immediately any desired image. The high-throughput HCS projects described by Dr. Leven were able to analyze 40,000 compounds on a daily basis, for a total campaign of more than two million compounds, generating multifeatured data sets for each well.

PerkinElmer’s high-content screening portfolio includes the Opera confocal microplate image reader and Acapella™ image-analysis software, the compact Operetta HCS system, driven by Harmony™ software, and the Columbus™ data-management system and new Columbus 2.0 for use with the Opera platform.

Gabriele Gradl, Ph.D., global product leader for HCS at PerkinElmer Cellular Technologies, emphasized the complexity involved in deriving robust, quantitative data from cellular measurements derived on image analysis of high-content screens. Whereas, fluorescence-based analysis typically relies on identifying objects in cells and measuring their fluorescence intensities, PerkinElmer has developed a computational strategy that is independent of absolute fluorescence intensity. It relies on texture analysis and quantitative pattern analysis for data generation.

Texture-analysis tools can detect patterns and effects that would not be apparent on routine visual analysis, according to Dr. Gradl. Threshold adjacency statistics is one example of such a tool. It searches for differences in fluorescence intensity values between adjacent pixels over a defined distance. Dr. Gradl described the particular advantages of applying texture analysis for detecting subtle morphologic changes associated with cell viability or toxicity assays and in stem cell research. It can detect differences not visible to the eye and identify changes that the user might not even have known to look for in the data. She presented, as an example, the use of texture analysis to assess mitochondrial integrity, as loss of mitochondrial activity and enhanced mitochondrial biogenesis are early markers of cytotoxicity.

Dr. Gradl also described the use of texture analysis in brightfield imaging and the ability to assess segmentation based on granularity, enabling label-free proliferation assays and analysis of cell differentiation in real time.

The algorithms developed by PerkinElmer can apply texture analysis to whole cells or to specific intracellular compartments depending on the design of the assay. The company is exploring a range of applications for its texture-analysis software tools, including stem cell differentiation analysis, quality control of stem cells produced for therapeutic use, live-cell imaging over time, and 3-D tissue sample analysis.

Earlier this year, GE Healthcare introduced the IN Cell Analyzer 2000 cell-imaging system, which incorporates several new features: preview scoring of a selected area of a sample before an acquisition run; a large chip CCD camera coupled with a widefield illumination source for twice the brightness of a conventional xenon lamp, according to GE; whole-well imaging; an objectives range from 2x–100x; six imaging restoration modes; and a manual microscope mode.

Fred Koller, Ph.D., president and CEO of Cyntellect, launched the company’s new Celigo™ cytometer at the “High Content East” meeting, emphasizing the system’s ability to image “every cell in every well,”  from edge to edge without edge effects using both brightfield and fluorescence imaging. Cyntellect’s optical technology achieves high-quality large field imaging using a set of mirrors to capture each well in its entirety without moving the plate and without the need to refocus, allowing for rapid, full-plate imaging.

Celigo provides “uniform illumination with no gradient across the well,” said Dr. Koller, and allows for a combination of label-free imaging and three-color fluorescence. He described the instrument’s capabilities for performing cell-counting assays, cell growth tracking, and confluency studies, for example, and for noninvasive imaging of stem cell cultures without disrupting their three-dimensional colony structures. Celigo can switch from single-cell to colony-counting mode.

The company has also developed a secretion assay for use on the Celigo that measures the amount of protein secreted by individual cells. The assay can distinguish between high and low secretors and is useful for detecting heterogeneity and instability in cell cultures early in process development, such as for antibody manufacturing.

The Cellular Imaging and Analysis group at Thermo Fisher Scientific introduced the Cellomics iDev™ intelligent assay development workflow for HCS image analysis at “High Content East”. Users work training image sets of positive and negative biology, applying imaging and analytical algorithms that allow for real-time interaction with the images. The software employs the biological data generated to optimize assay protocols.

Source: genengnews.com

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Researchers find candidates for new HIV drugs

Last Updated on Tuesday, 1 December 2009 11:26 Written by Editor Tuesday, 1 December 2009 11:26

While studying an HIV protein that plays an essential role in AIDS progression, researchers at the University of Pittsburgh School of Medicine have discovered compounds that show promise as novel treatments for the disease.

HIV drug discovery efforts have met with little success in finding compounds that interact with an important HIV virulence factor, called Nef, because it lacks biochemical activity that can be directly measured, explained Thomas E. Smithgall, Ph.D., William S. McEllroy Professor and Chair, Department of Microbiology and Molecular Genetics, and senior author of the paper, which was published last week in the early, online version of ACS Chemical Biology.

To get around that problem, Dr. Smithgall’s team developed an assay to measure Nef function indirectly by coupling it to another protein, called Hck, which Nef activates in HIV-infected cells. Because Hck activity can be easily measured, the investigators were able to use it as a reporter for Nef activity in an automated high-throughput screening process. In collaboration with the University of Pittsburgh Drug Discovery Institute, they screened a library of 10,000 chemical compounds against the coupled proteins to see which ones influenced Nef-induced activation of Hck.

After further testing, they confirmed that three compounds inhibited the activity of the Nef-Hck complex and, more importantly, all of them also interfered with HIV replication. One compound was so effective that it suppressed HIV replication to undetectable levels in cell culture experiments.

“So we now have a way to rapidly and efficiently screen for inhibitors of Nef signaling through Hck,” Dr. Smithgall said. “But the surprise was that some of those inhibitors also showed strong antiviral activity in cell culture models.”

There is evidence that people infected with HIV variants that have mutations in the Nef gene take substantially longer to develop disease symptoms or AIDS, he said. In animal models, disrupting the production of Nef from the virus or its interaction with Hck also delays or prevents disease symptoms. The next challenge for the researchers will be to determine whether these compounds also interfere with progression of AIDS-like disease in animal models by blocking Nef function.

“Most current therapies for HIV infection use drugs that interfere with the function of viral enzymes such as reverse transcriptase or with the interaction of the virus and the host cell,” Dr. Smithgall said. “Targeting Nef represents an entirely new approach that could be useful to deal with issues such as drug-resistant HIV strains, and may slow the progression to AIDS.”

He added that Nef is just one of several so-called “accessory proteins” encoded by HIV which are important virulence factors in AIDS. Inhibitory compounds against some of the others might be revealed using a similar coupled protein approach for high throughput screening.

Source: labspaces.net

Posted under Compound Screening, Discoveries, Innovations and Patents, Drug Development, HIV Research, HT Screening, New Drugs, Press Releases  |  Comments Off

MALDI-Based Method May Reduce Cost of Rx-Screening Assays, Speed Drug Development

Last Updated on Tuesday, 20 October 2009 09:44 Written by Editor Tuesday, 20 October 2009 09:44

This story originally ran on Oct. 6.

By Tony Fong

Researchers from the University of Cincinnati and MDS Analytical Technologies have used mass spectrometry to develop a high-throughput screening method for drug discovery they say can be more precise and cost-effective than existing techniques.

The technique is based on a MALDI triple-quadrupole platform and exploits the selective multiple-reaction monitoring transition features of the platform. By doing so, the new method is able to lower the cost of high-throughput screening for drug compounds to pennies per well from as much as $1 per well currently, Ken Greis, associate professor of cancer and cell biology and director of proteomics and mass spectrometry at the University of Cincinnati College of Medicine, told ProteoMonitor recently.

A study detailing the method was published Sep. 15 in the online edition of Rapid Communications in Mass Spectrometry.

In the paper, Greis and his co-authors said that drug discovery typically begins with a validated target enzyme “with the initial goal of finding appropriate molecular scaffolds with inhibitory activity via high-throughput screening.” The scaffolds are then subsequently used for lead compound optimization, and “ideally for the development of a safe and effective therapeutic compound.”

The most common methods of high-throughput screening have been fluorescence- and chemiluminescence-based approaches. Such approaches, Greis said, have been “very successful” because the same reagents can be used for many different enzymes.

But that same characteristic also creates a risk for interference.

“When one’s evaluating a compound repository for inhibitors, you often have a series of compounds that will fluoresce themselves,” Greis said. “If they fluoresce, they’re going to give you a false signal. Alternatively … there are compounds that inhibit the fluorescent properties, or what’s called quenching fluorescence, [that] also give false read-outs.”

Another problem is in the way the assays get generalized so that the reagents work for a wide range of enzymes. Such assays are called coupled assays: “You have a product being formed from your enzyme reaction but that’s not what actually triggers the fluorescence,” Greis said. “That product gets converted to another enzyme to another product through another enzyme to another product that then can be fluoresced.”

This series of enzyme step, or coupled assays, ultimately results in a read-out. “The problem is any compound that interferes with any of those steps along the way also gives you false read-outs,” which tend to be false positives, he added.

But by using mass spectrometry to measure enzyme activity, Greis and his colleagues are able to get a direct read-out, “so a mass spectrometer effectively can give you a quantitation and a mass of a compound.”

By taking a ratio of the substrate being converted to a product — the essence of an enzyme assay, Greis said — and measuring that directly on a mass spec, there is no interference either from quenching or auto-fluorescence.

“And what we’ve found thus far is we’ve not seen any false positive read-outs. If we get a compound that shows that it’s active, even in single-point assays, it’s been demonstrated that it’s a dose-dependent inhibitor.”

And because the method uses native peptides or small-molecule substrates, the method can be done for “at most, pennies per sample well,” Greis said. By comparison, fluorescent and chemiluminescence reagents cost between 50 cents to $1 per well.

“So if you run a million compounds, you can run up a half-million dollars of reagents costs right away, whereas the label-free read-out is going to cost you maybe a couple thousand dollars for the reagents,” he said. “That’s a mass spec advantage.”

A prior study by researchers in China had demonstrated the utility of a MALDI-Fourier transform mass spectrometer for high-throughput screening of small-molecule substrate/product conversion.

In their work, though, Greis and his co-researchers wanted to extend the application to a small-molecule, non-peptide substrate to demonstrate the flexibility and technical range of their method. While a large number of therapeutic targets, including kinases and phosphatases, contain peptide substrates, some important targets don’t, such as HMG CoA reductase, the target of statin drugs, and AChE, a target for neurodegenerative therapeutics.

The researchers chose AChE because of its long history of enzyme assay development, including colorimetric assays, pH-change assays, and most recently aggregation-induced fluorescence assays and mass-spec assays.

Speed is of the Essence

They also chose a MALDI platform, rather than an electrospray platform, because of the higher speed that can be achieved on the MALDI. Most enzymatic reactions contain salts that can interfere with mass spectrometry. An ESI platform requires a desalting step, which limits the throughput to five to 10 seconds per sample. A MALDI-based approach skips the desalting step, however, because the technology is less sensitive to salts.

“Essentially all that we do is run the enzyme reaction on a 384-well format,” Greis said. “We transfer all at once into a matrix plate mix and onto our MALDI target plate.”

Because there are no cleanup steps on the MALDI triple-quad, samples can be scanned at up to three samples per second, he said.

Greis acknowledged that the MALDI technology, especially the MALDI triple-quad, is not a popular tool for drug discovery. In his opinion, that’s because drug-discovery researchers were trained on electrospray mass specs and are comfortable with them.

“To then move them into a MALDI platform that they don’t understand, they’ve got a bias that it can’t be quantitative, and all these sorts of things from earlier studies using MALDI-based approaches that have been demonstrated time and time again to not be true anymore — I think there’s a cultural thing,” he said.

A criticism of a MALDI approach is that while it works well for peptide substrate screening, it doesn’t for small-molecule substrate products because of matrix interference in the low mass range.

“And we show very directly … that by taking advantage of the transition,” a chemical fragmentation that is diagnostic of a substrate or product “that one can do in a triple-quad, that matrix interference completely goes away,” Greis said.

The researchers tested their method by screening a library of 1,008 structurally diverse compounds across 384-well microtiter plates as an example of a single-dose primary screen, and reported that all known AChE inhibitors resulted in complete inhibition of enzyme activity, as expected. The hits were then validated “by demonstrating concentration-dependent inhibition and the rank order of inhibitory potency in hit follow-up assays,” they said in their study.

The technique they’ve developed can also be used on a simple MALDI instrument, though it works best for peptide substrate enzymes. With low molecular-weight enzymes, sensitivity can be an order of magnitude lower on a simple MALDI “because you’d have to be using enough enzyme substrate product to see your substrate products down in those low mass ranges in amongst all of the matrix peaks,” Greis said.

Also, Greis said there will be enzymes — such as fatty acids and long-chain hydrocarbons —that will not be amenable to a MALDI-based approach.

“The fact of the matter is that any mass spectrometry-based technique is only as good as the molecule that it’s trying to evaluate,” he said. “We have to be able to ionize the substrate and/or the product to be able to measure and quantify it.”

In ongoing work, he and his team members are developing multiplex assays. The typical screening approach is to take a target enzyme and pass the whole repository across it to look for inhibitors, and then validate the inhibitors. The next therapeutic target is then set up and the process is repeated.

With a mass spec-based approach, “as long as your enzymes reactions are compatible … you can run multiple enzymes in one pot and pass your repository against it once and get hits for all those different enzymes,” Greis said.

In conferences, Greis and his colleagues have presented proof-of-concept studies that show that “this in fact works quite well using a kinase and acetylcholinesterase or a kinase with a protease all in the same part,” he said. “We’ve shown that we can get selective inhibitors for each of them individually without interference in the multiplex format.”

Source: genomeweb.com

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Structure-based substrate screening for an enzyme

Last Updated on Wednesday, 26 August 2009 11:20 Written by Editor Wednesday, 26 August 2009 11:20

Nowadays, more and more novel enzymes can be easily found in the whole enzyme pool with the rapid development of genetic operation. However, experimental work for substrate screening of a new enzyme is laborious, time consuming and costly.

On the other hand, many computational methods have been widely used in lead screening of drug design. Seeing that the ligand-target protein system in drug design and the substrate-enzyme system in enzyme applications share the similar molecular recognition mechanism, we aim to fulfill the goal of substrate screening by in silico means in the present study.

Results: A computer-aided substrate screening (CASS) system which was based on the enzyme structure was designed and employed successfully to help screen substrates of Candida antarctica lipase B (CALB).

In this system, restricted molecular docking which was derived from the mechanism of the enzyme was applied to predict the energetically favorable poses of substrate-enzyme complexes. Thereafter, substrate conformation, distance between the oxygen atom of the alcohol part of the ester (in some compounds, this oxygen atom was replaced by nitrogen atom of the amine part of acid amine or sulfur atom of the thioester) and the hydrogen atom of imidazole of His224, distance between the carbon atom of the carbonyl group of the compound and the oxygen atom of hydroxyl group of Ser105 were used sequentially as the criteria to screen the binding poses.

223 out of 233 compounds were identified correctly for the enzyme by this screening system. Such high accuracy guaranteed the feasibility and reliability of the CASS system.

Conclusions: The idea of computer-aided substrate screening is a creative combination of computational skills and enzymology.

Although the case studied in this paper is tentative, high accuracy of the CASS system sheds light on the field of computer-aided substrate screening.

Author: Tao XuLujia ZhangXuedong WangDongzhi WeiTianbi Li
Credits/Source: BMC Bioinformatics 2009, 10:257

Source: 7thspace.com

Posted under Compound Screening, Discoveries, Innovations and Patents, HT Screening, Press Releases  |  Comments Off

GTCbio Announces 4th Annual Assay Development and Screening Conference taking place June 8-9, 2009.

Last Updated on Friday, 27 March 2009 09:40 Written by admin Friday, 27 March 2009 09:40

San Francisco, CA – GTCbio Announces its 4th Annual Assay Development and Screening Conference taking place June 8-9, 2009. As compounds derived from high throughput screening increasingly find their way into clinical trials, drug screening has become widely accepted as a critical step in the drug discovery process. After more than a decade of rapid growth, tremendous progress has been made in assay technology, laboratory automation, and informatics. These technological developments have not only facilitated a drastic increase in throughput and efficiency in drug screening, but have also provided novel solutions in other areas of drug discovery and development. As screening has also become prominent in biological research, screening facilities have become increasingly popular in academic institutions.

As the pharmaceutical industry continues to face the challenges of developing more new chemical entities and reducing the cost of R&D, the demand for novel technologies and creative approaches for improving the efficiency of screening has intensified. Cell-based assays used in compound screening and high-content screening technologies have gained popularity in the industry. Years of intensive research have finally resulted in label-free technologies in the drug screening market place. These technologies provide new ways of interrogating cellular and molecular binding events and enable orthogonal screening approaches to drug targets.

The goal of the 4th annual Assay and Screening Technologies Conference is to provide a forum for academics and professionals in the drug discovery industry to stay abreast of exciting new developments in assay technologies while exchanging ideas and developing more efficient approaches to the drug discovery and development process.

For more information, visit http://gtcbio.com/conferenceDetails.aspx?id=123

Posted under Clinical Trials, Compound Screening, Drug Development, HT Screening, North America, Press Releases, USA and Canada  |  Comments Off

Fragment Based Screening Service at CRELUX and ZoBio

Last Updated on Thursday, 27 November 2008 10:04 Written by admin Thursday, 27 November 2008 10:04

Munich (D) and Leiden (NL), November 24, 2008 / b3c newswire /  – CRELUX and ZoBio announced today that they have successfully executed their first fragment based screening projects from a jointly established platform.

One of the first targets, which also will be made accessible to customers, was Pim1, a kinase that has been implicated in the progression of several haematological malignancies. In addition to the “off the shelf” data on Pim1, tailor made fragment based screening projects are available for customers upon request.

The joint technology platform combines ZoBio’s proprietary Target Immobilized NMR Screening (TINS) technology with CRELUX’s high performance kinase crystallography platform. In the first campaign Pim1 was screened by TINS using ZoBio’s fragment library, hits were assessed in an in vitro kinase assay and the top 50 hits have been soaked into protein crystals. 37 out of these 50 fragments showed defined binding modes. Together with this high hit rate the structural diversity within this group generated multiple points for optimization and clearly proved the power of this technology combination.

“We are delighted to have found a perfect partner for entering into high performance fragment based screening. The collaboration with ZoBio adds another crucial drug discovery technology to our service portfolio”, commented Dr. Michael Schäffer, CEO of CRELUX.

“This project demonstrates the power of the combination of TINS with top notch crystallography. I am absolutely convinced that together we can provide our customers with critical starting point to jump start their challenging or failed targets.” noted Dr. Gregg Siegal, CSO of ZoBio.

CRELUX has used its state-of-the-art structural biology platform to solve more than 270 crystal and co-crystal structures for pharma and biotech companies. This platform encompasses all steps – from target cloning and expression all the way to high-throughput protein crystallization and in-house x-ray crystallography.

ZoBio provides fragment discovery and characterization services to the pharmaceutical and biotech industries using its proprietary Target Immobilized NMR Screening (TINS) platform. TINS, with its unparalleled sensitivity and reliability, has been used to discovery highly diverse, efficient ligands for a variety of targets including kinases, protein-protein interactions, viral targets and membrane proteins.

Posted under Collaborations, Diversity Libraries, Europe, HT Screening, New Products, Press Releases, Targeted Libraries  |  Comments Off

Cell-Based Assays: Innovations in Reagents, Technologies & Screening October 23 – 24, 2008

Last Updated on Thursday, 23 October 2008 12:56 Written by admin Wednesday, 22 October 2008 04:56

Cell-based assays provide one of the most valuable tools in drug discovery.  They are routinely used in target validation, HTS campaigns, structure activity relationship analyses and ADMET studies. Many factors need to be considered in designing, developing and running relevant cell based assays to progress discovery programs.  Significant advances continue to be made in assay design and cell supply processes that incorporate biologically relevant cell types and novel detection technologies. This symposium is designed to capture successful techniques and practices that enable high quality cell-based assays using commonly employed cell types such as CHO, HEK and U-2 OS, as well as the expanding application of additional cell types. The symposium will be of great interest to cell culture scientists, assay developers, screeners, medicinal chemists, ADMET and therapeutic teams.

Session Overviews:

Session 1: Cells as Reagents – Fact or Fiction?
This session will cover various aspects of the most important material used in cell-based assays: the cells. The talks will address the topics such as: how to characterize cell lines to ensure their identity? What are the impacts to the cells after transfection? What are the issues during scale up process? This session will also highlight advances in cell culture automation and material tracking system for cell-based assays.

Session 2: Assay Development – Present Realities
Topics covered include the application of BacMam virus based gene delivery in assay development, the development of high-content cellular assays, dielectric spectroscopy technology and photoprotein aqueorin based GPCR assay platforms and considerations in screening for antibody based therapeutics.

Session 3: Cell-Based Screening – The How, Why & Where
This session will include topics on the implementation of cell-based assays and screens across the discovery process focusing on the challenges, solutions and issues to consider. Presentations include experiences with high throughput screening for lead identification, profiling with cellular panels, ADME applications, and the use of high content screening in both drug discovery and to probe complex cellular systems.

Session 4: Cell-Based Assays – Emerging Trends
The topics in this session will address emerging trends in supply of cells that attempts to bridge the gap between traditional target based in vitro assays and in vivo measurements. The presentations will highlight potential applications of cells to build model systems that can offer the combined benefits of traditional in vitro and ex vivo approaches. These emerging technologies and methodologies hold the promise of addressing a major gap in using target based approaches to discover new biological tools and drugs and will challenge the supply of cell reagents.

http://www.sbsonline.com/

Posted under ADMET Studies, Cell-based Assays, Genomics & Pharmacogenomics, HT Screening, Medicinal Chemistry, North America, Press Releases, USA and Canada  |  Comments Off

Sanofi-aventis signs a collaboration agreement with RainDance Technologies and Louis Pasteur Univers

Last Updated on Wednesday, 1 October 2008 03:48 Written by Fred Wednesday, 1 October 2008 03:48

The FINANCIAL — Sanofi-aventis announced on September 26 the launch of the dScreen Consortium, a research initiative conducted with RainDance Technologies, Inc., Lexington, Massachusetts, and Louis Pasteur University, Strasbourg, France, to develop the next generation of High-Throughput Screening (HTS) for drug discovery applications.

The consortium was founded with the assistance of the Alsace BioValleyTM Cluster, France, which helped secure financing and support for the program.
The dScreen Consortium assembles:
- the renowned drug screening expertise of sanofi-aventis
- the unique expertise in droplet-based microreactors of the Chemical Biology Laboratory at the Institute for Science and Supramolecular Engineering (ISIS) of Louis Pasteur University
- and RainDance Technologies’ unique capabilities to apply droplet-based microfluidic technologies to human health and disease research.
“We are delighted to enter this partnership with two highly innovative research groups in this rapidly advancing field,” said Martin Galvan Ph.D., Scientific Director at the sanofi-aventis research site in Strasbourg. “The expected gains in terms of productivity and knowledge should significantly accelerate our drug discovery programs”.
Based in the Alsace BioValley in Strasbourg, the consortium will utilize the pico-liter volumes and ultra-high speed capabilities of RainDance’s technology and systems to achieve breakthrough performance in high-throughput drug screening methodologies.
“This exciting project represents the first research collaboration undertaken by our new RainDance Technologies France SARL subsidiary,” said Chris McNary, President and Chief Executive Officer of RainDance Technologies. “The speed, simplicity, and minute volume of our droplets eliminate the need for unnecessarily complex automation solutions in high-throughput screening. Our technology will process 10 million droplets per hour on a single benchtop instrument, dramatically accelerating the drug discovery process while conserving precious screening compounds,” added McNary.

“This project is an excellent opportunity to develop the compartmentalisation of reactions in emulsion droplets for an entirely new field of application: HTS for drug discovery” said Andrew Griffiths, head of the Chemical Biology Laboratory at ISIS. ”University Louis Pasteur is proud to be part of this consortium which will open new scientific routes while generating top scientific lectures to our students” said Jean-Marc Jeltsch, Vice-President, Louis Pasteur University.
“The dScreen Consortium is a great illustration of the “Pôle de Compétitivité” policy in France: the development of breakthrough innovations in drug screening through collaborative R&D programs results in strengthening local actors such as the sanofi-aventis research site and in the creation of a subsidiary of an US company in Alsace. Furthermore, the establishment of a drug screening services platform based on the results of the program will reinforce the capabilities of our cluster” underlined Pascal Neuville, President of Alsace BioValley.

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Idealp-Pharma launches « hit-to-candidate » services

Last Updated on Friday, 18 July 2008 06:36 Written by admin Friday, 18 July 2008 06:36

Services to accelerate programs from biological target to first-in-man use Idealp-Pharma is launching fully integrated drug discovery and preclinical development services combining medicinal chemistry, cheminformatics,
screening, early ADMET and preclinical development capabilities to speed up
partner’s and client’s small molecules programs from biological target to firstin-
man use.

According to Serge Petit, PhD, President and CEO, “Being a one-stop-shop company adds significant value because the lead optimisation process involves iterative cycles for incremental optimization. The main advantages of our one-stop-shop service are to have access to all the experimental data, to be able to refocus the synthesis program and then to make the best decision for the lead optimisation process in accordance with our customers’ specifications.”

“Idealp-Pharma manages its customers’ hit discovery and validation, hit-to-lead
progression and lead-to-candidate process. Our aim is to deliver chemically and
biologically validated hits, accelerating lead optimization and identying IND candidate for our customers”, said Serge Petit. Idealp-Pharma supports also its client’s drug discovery activities by providing modular and customized services such as medicinal chemistry and cheminformatics studies.

More information about integrated drug discovery services can be found at www.idealp-pharma.com
About Idealp-Pharma

Idealp-Pharma’s aim is to expand partner’s drug pipeline by accelerating drug
discovery process from the biological target to first-in-man use. Idealp-Pharma
provides a range of flexible services: including fully integrated drug discovery and preclinical development, medicinal chemistry and cheminformatics.

Idealp-Pharma’s purpose-built lab covers a total of 2000 square meters. Idealp-Pharma now employs 60 staff. More information about Idealp-Pharma can be found at www.idealp-pharma.com

Posted under ChemInformatics, Drug-Like Compounds, Europe, HT Screening, Medicinal Chemistry, Press Releases  |  Comments Off

Genedata and Axxam Announce High Throughput Screening Data Analysis Collaboration

Last Updated on Wednesday, 9 July 2008 03:41 Written by Fred Wednesday, 9 July 2008 03:41

Genedata and Axxam have signed a multi-year contract for the implementation of the Genedata Screener® data analysis and management platform into Axxam’s high throughput screening process.

Genedata, the leading provider of in silico solutions for the pharmaceutical and life science industries, and Axxam, a leading biotechnology research organization offering early-stage discovery research services for the life science industry, have signed a multi-year contract for the implementation of the Genedata Screener® data analysis and management platform into Axxam’s high throughput screening process.

With an increasing number of pharma and life science companies outsourcing entire stages of their research cycle, the contributions of contract research organizations (CROs) to drug development have never been more crucial. Faster development, earlier decisions on project failures, and higher approval success rates are becoming the norm. To continuously meet those high standards, Axxam relies on Genedata Screener. Screener’s high-quality, high-performance modular system rapidly and flexibly analyzes, integrates and manages all assay data and then combines it with chemical, pharmacological and in vivo information to support Axxam scientists in prioritizing compounds and identifying quality lead structures with the highest confidence. Screener`s open and scalable architecture integrates with existing infrastructures, which allows it to be tailored to specific discovery processes, maximizing value.

Dr. Stefan Lohmer, CEO of Axxam, said, ”Establishing flexible and efficient working practices to ensure client satisfaction is our top priority as a reputable service provider. Genedata’s consistently high performance and data quality along with their expertise and profound scientific knowledge enable us to meet that goal on a daily basis, with rigorous quality control covering every detail, as well as keeping to deadlines and budget terms. Thus, our focus extends beyond the actual license agreement with Genedata, towards a comprehensive, long-term partnership in high throughput screening.”

Dr. Othmar Pfannes, CEO of Genedata, stated, “We are proud to have met Axxam’s high selection standards. Genedata as leading research informatics provider and Axxam as ‘best-in-class’ contract research organization for life science discovery research are creating a powerful synergy here that lets us look with excitement to the future.“

Axxam is a science-driven biotechnology company, which offers early-stage discovery research services for the life science industry. The company applies its comprehensive from genes to leads activities to Discovery Services that we provide for customers and to internal Discovery Research. Activities include assay development, high-throughput screening (HTS) and compound profiling. A team of almost 70 qualified personnel is committed to advance the discovery projects of our partners. The scientific know-how of our team is built upon years of experience as part of the assay development group of the Bayer HealthCare, Research and Development organization prior to the late 2001 inauguration of Axxam as an independent private company. The growing list of our collaborations in the life science industry is recognition of our success in areas such as the pharmaceutical, agrichemical, fragrance, cosmetic, flavour, food and beverage branches. Axxam is a privately-owned company with offices and state-of-the-art laboratories located in the San Raffaele Biomedical Science Park, Milan.

Genedata specializes in discovery informatics for biotech, pharmaceuticals and the life sciences. The company offers expertise in research informatics combined with open and scalable computational solutions. Our product suites include Genedata Phylosopher® for integrating, structuring, and analyzing research data, Genedata Screener® for high throughput screening, high content screening analysis and hit-to-lead, and Genedata Expressionist® for omics data integration, processing and analysis. Founded in 1997 as a privately held spin-off from Novartis, Genedata is headquartered in Basel, Switzerland, and has branches in Munich (Germany), Konstanz (Germany), Boston (USA), San Francisco (USA), and Tokyo (Japan). The company is also represented in Taiwan and Singapore.

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4th Annual International Conference on Compound Libraries – in Dusseldorf, Germany from 6th to 8th October 2008.

Last Updated on Tuesday, 1 July 2008 01:46 Written by admin Tuesday, 1 July 2008 01:46

From Focused Compound Libraries to optimised Hit-to-Lead – that’s the motto of IQPC’s 4th international conference on “Compound Libraries” (formerly “Focused Compound Libraries”). Nowadays the pharmaceutical industry is under enormous pressure, and the key for the industry to survive is faster and more efficient drug discovery. To improve their lead generation process and library, pharmaceutical companies need to choose the correct library design from the very beginning. Also, they need to integrate new compounds and collections into their library design to guarantee its continuous improvement. However, urgent questions still remain: How can you find the ideal library size to assure diversity while keeping focused? Can fragment based screening speed up the discovery process? How can you guarantee the best screening outcome analysis to ensure lead optimization?

After concentrating on focused compound libraries in the past years, this year presentations will cover the design and enhancement of different kinds of libraries as well as the possibilities of hit-to-lead optimization.

Maximize your knowledge of the latest advances in intelligent library design:

  • Explore how to efficiently integrate new compounds and collections to improve the lead generation process
  • Learn how to build up an effective collection of compounds in your company to guarantee physical quality and quantity of the compounds
  • Hear about enhanced screening methods such as fragment-based screening to reduce complexity in the screens
  • Successfully enlarge your compound collection by utilising novel structures and multi-component reactions in library design
  • Enhance your Hit-to-Lead ratio through advances in library design, screening methods and structure based drug discovery approaches

Experts from international companies such as Pfizer, AstraZeneca, Merck, Grünenthal, Sanofi-Aventis and many more will report about first-hand experiences and best practices.

Full speaker line-up:

  • Sanofi-Aventis Gruppe, Germany
  • AstraZeneca Ltd., UK
  • Merck Serono, Germany
  • Basilea Pharmaceutica International AG, Switzerland
  • Organon Laboratories Ltd., UK
  • GenKyotex S.A., Switzerland
  • Carlsberg Laboratory, Denmark
  • Chemical Genomics Centre of the Max-Planck-Society, Germany
  • AstraZeneca, R&D Mölndal, Sweden
  • AnalytiCon Discovery GmbH, Germany
  • Asinex Ltd., Russia
  • TU Vienna, Austria
  • BioFocus DPI Limited, UK
  • Solvay Pharmaceuticals BV, Netherlands
  • Grünenthal GmbH, Germany
  • Novartis, Switzerland
  • Bayer CropScience AG, Germany
  • Pfizer Ltd., UK

http://www.iqpc.com/ShowEvent.aspx?id=113724

Posted under Compound Libraries, Compound Screening, Diversity Libraries, Drug-Like Compounds, Europe, Europe, HT Screening, Medicinal Chemistry, Press Releases  |  Comments Off

Direct Screening of Natural Products Extracts Using Mass Spectrometry

Last Updated on Monday, 30 June 2008 04:20 Written by admin Monday, 30 June 2008 04:20

Present advances in screening and separation technologies reveal extracts bioactivity with great efficiency and accuracy. Recently reported method of Direct Screening of Natural Products Extracts Using Mass Spectrometry does not require any preparation or fractionation work. Several hundred crude extracts can be screened per one day. Direct bioaffinity screening mass spectrometry method followed by the use of ligand mass information for mass-directed purification makes screen of crude extracts and identification of active compounds precisely possible.

Vu, H., et. al. Direct Screening of Natural Product Extracts Using Mass Spectrometry. J. of Biomol. Screening. 2008, v. 13, 4, 265-275

Plants and plant extracts from Timtec

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Spermatech Selects Evotec as Partner for High-Throughput Screening and Lead Discovery

Last Updated on Tuesday, 18 March 2008 03:34 Written by Fred Tuesday, 18 March 2008 03:34

HAMBURG, Germany and OXFORD, England and OSLO, March 12, 2008 /PRNewswire-FirstCall/ — Evotec AG announced today that Spermatech A/S has chosen them as a partner to identify small molecule therapeutics for their pharmaceutical discovery project.

Through the study of the physiology of sperm motility, more specifically of “rapid swimmers” that cause fertilisation, Spermatech have identified biological targets that could be exploited in the development of non-hormonal reversible male contraceptives. On this basis, Evotec and Spermatech have defined a strategy for a tailored drug discovery project. Evotec will apply its expertise and proprietary technologies in assay development, high throughput screening and NMR (Nuclear Magnetic Resonance) screening to identify inhibitors of the sperm specific target protein. The screening will be performed with Evotec’s screening library of 250,000 drug-like compounds. Compounds will be identified that reduce sperm motility and will be used in the development of non-hormonal reversible male contraceptives at Spermatech. In addition, compounds that promote target activity may be evaluated as supporters of male fertility.

Dr Mark Ashton, Executive Vice President Business Development Services at Evotec, said: “We are extremely pleased that Spermatech has selected Evotec for this interesting project. It will allow us to use our combined technologies in assay development, high-throughput screening and NMR screening to identify the most promising candidates in the therapeutic field. Evotec’s highly diverse compound library is a good starting point to identify such active molecules and the additional results from NMR investigations of the hits with the target protein will provide the medicinal chemists with useful information to support subsequent drug design.”

“We were impressed by Evotec’s highly specialized and integrated capabilities. The collaboration will provide us with access to state-of-the-art assay development and screening technology and expertise together with a high quality library of small molecules. We are confident that this will provide an excellent starting point and valuable information to progress the molecules into more advanced stages. We really appreciated that during the initial scientific discussions of the project Evotec clearly demonstrated a results-oriented spirit in support of our project.” commented Eirik Naess-Ulseth, Chief Executive Officer, Spermatech.

Forward looking statements

Information set forth in this report contains forward-looking statements, which involve a number of risks and uncertainties. Such forward-looking statements include, but are not limited to, statements about the anticipated benefits of Evotec’s products and services, the payments that Evotec may receive under its collaboration agreement with Spermatech, the anticipated timing and results of Evotec’s clinical and pre-clinical programs, and other statements that are not historical facts. Evotec cautions readers that any forward-looking information is not a guarantee of future performance and that actual results could differ materially from those contained in the forward-looking information as a result of risks and uncertainties. These include risks and uncertainties relating to: Evotec’s ability to complete the merger because conditions to the closing of the merger may not be satisfied; the failure to successfully integrate the businesses of Evotec and Renovis; unexpected costs or liabilities resulting from the merger; the risk that synergies from the merger may not be fully realized or may take longer to realize than expected; disruption from the merger making it more difficult to maintain relationships with customers, employees or suppliers; competition and its effect on pricing, spending, third-party relationships and revenues; the need to develop new products and adapt to significant technological change; implementation of strategies for improving internal growth; development, use and protection of intellectual property; general worldwide economic conditions and related uncertainties; future legislative, regulatory, or tax changes as well as other economic, business and/or competitive factors; and the effect of exchange rate fluctuations on international operations.

The risks included above are not exhaustive. The Registration Statement on Form F-4 filed by Evotec with the Securities and Exchange Commission contains additional factors that could impact the combined company’s businesses and financial performance. The parties expressly disclaim any obligation or undertaking to release publicly any updates or revisions to any such statements to reflect any change in the parties’ expectations or any change in events, conditions or circumstances on which any such statement is based.

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US Plans Use of Automated Toxicity Screening, Could End Animal Testing

Last Updated on Wednesday, 27 February 2008 05:10 Written by Fred Wednesday, 27 February 2008 05:10

Federal scientists are collaborating on a new approach to testing the toxicity of chemicals ranging from pesticides to household cleaners. They plan to use new automated high-speed cell tests to get more reliable data faster and more cheaply, and with less reliance on animal testing.

The National Institutes of Health and the U.S. Environmental Protection Agency are partners in the plan. NIH director Elias Zerhouni says scientists will apply technology developed by the two agencies to identify chemicals that might be harmful to humans.

“You could, in a battery of tests, end up with very specific molecular signatures that will be predictive of human toxicology, in ways that you just can’t do in animal testing today,” he said.

Scientists now rely heavily on animal tests to generate chemical toxicity data. That process is expensive, time-consuming and not always the best predictor of effects on humans says Francis Collins, director of the NIH National Human Genome Research Institute.

“There are differences between species. We are not rats and we are not even other primates, and so that [the] desire here is to see if we could do better,” she said. “Ultimately what you are looking for is [whether] this compound does damage to cells.”

The high-speed automated screening looks at the effects of chemical compounds on single human cells rather than on an entire laboratory animal. Researchers expect the new toxicology testing method will expand the number of chemicals tested and reduce the time, money and use of animals. NIH director Elias Zerhouni says it will also generate data more relevant to humans.

“What is being proposed here is to move the 20th century paradigm of testing of one compound at a time in many animals to going to the 21st Century paradigm [that] tests 5,000 to 10,000 compounds against 20,000 conditions in cells that are very specific to human toxicology,” he said.

Since NIH started the National Toxicology Program 30 years ago, it has tested 2,500 chemicals. Using the new automated strategy could get the same job done in a single afternoon.

John Butcher, associate director of the National Toxicology Program, explains that to check the reliability of this approach, scientists will first do a comparative analysis of the 2,500 chemicals previously tested on animals.

“And we can compare the output from these cell-based assays, in terms of whether these chemicals cause cancer, reproductive and developmental effects, neurological effects, immunotoxic effects and various other kinds of toxicity,” he said.

Butcher says the anticipated shift away from animal testing could take many years. Writing in Science Magazine, he says the agencies expect broader participation from public and private partners in the scientific community as the cell-based testing methods are refined and accepted.

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High Throughput Screening 2007: HighTech Business Decisions Reports New Strategies,

Last Updated on Thursday, 13 December 2007 05:02 Written by Fred Thursday, 13 December 2007 05:02 

SAN JOSE, Calif.--(Business Wire)--HTS laboratories continue to evolve their processes and strategies
to increase their success rates and improve efficiencies in their drug
discovery efforts. New strategies include the use of phenotypic and
pathway-based assays in primary or secondary screening, the use of new
targets and the expansion in the diversity of compounds screened for
new drug candidate leads. The Directors at HTS laboratories also plan
to employ new technologies, including the use of label-free
technologies and new detection systems to aid in finding new drug
candidate leads.

HTS laboratories continue to show year to year success at
producing new drug candidates. Fifty-five directors from HTS
laboratories participating in this recently published study, High
Throughput Screening 2007: New Strategies, Success Rates and Use of
Enabling Technologies, identified 163 drug candidates that originated
from their HTS laboratories, a significant increase from HighTech
Business Decisions' earlier study. With new HTS laboratories funded
through NIH or other private foundations beginning to ramp-up their
operations, this new report includes analysis and data from interviews
with HTS laboratory directors at these centers to better understand
their plans and their impact on drug discovery efforts.

William Downey, President of HighTech Business Decisions,
explains, "HTS laboratories are taking on new challenges as they
expand their role in drug discovery. In the past two years, the
industry has seen consolidation in HTS laboratories as many
pharmaceutical companies have either merged or acquired other
operations. This consolidation along with the challenges to improve
drug discovery efforts is leading to changes in the way particular HTS
laboratories operate. In addition to the changes in commercial HTS
laboratories, over the past two years publicly funded and
non-commercial HTS laboratories have been started, and in the next few
years these laboratories will ramp-up their operations." HighTech
Business Decisions extensively interviewed 55 Directors at HTS
laboratories in Asia, Europe and North America.

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Burnham Institute Selects Labcyte Acoustic Liquid Handling Technology

Last Updated on Saturday, 1 December 2007 08:06 Written by admin Saturday, 1 December 2007 08:06

Sunnyvale, CA November 15, 2007 – Labcyte Inc. will supply Burnham Institute for Medical Research (Burnham) with more than $2.3 million worth of Echo® liquid handlers for use in screening large biological libraries. The systems will be installed at both the Institute’s LaJolla, (CA) and Lake Nona, Orlando (FL) campuses. These instruments will be part of the world’s largest screening system and will bolster Burnham’s research and drug discovery efforts, which include research on cancer, neurological diseases, aging, diabetes, and obesity.

Burnham is one of the nation’s 10 collaborating centers of excellence funded under a special initiative created by the National Institutes of Health, which together comprise the largest public-sector drug discovery effort in history. The state-of-the-art Labcyte technology will improve screening results while reducing operating costs for Burnham and the collaborating network advancing this historical effort. The systems will be delivered over the next six months.

“The Echo liquid handlers were selected because their unique acoustic technology will enhance our ability to miniaturize assays, reduce compound waste, and meet throughput requirements,” explained Dr. Stefan Vasile, Director of the Chemical Library Screening Facility for Burnham Institute. “The Echo systems will be used for compound plate-to-plate transfer, reformatting, dose-response experiments and hit picking.”

“Our patented acoustic droplet ejection technology dramatically reduces waste while cutting operating expenses,” said Dr. Elaine J. Heron, Labcyte CEO. “Traditional liquid handlers are prone to poor precision and accuracy as transfer volumes are decreased. Researchers who require flexibility in their processes and biology find that the Echo systems provide that for them easily, quickly, and cost-effectively.”

Labcyte Inc., headquartered in Sunnyvale, California, is the world leader in providing acoustic droplet ejection technology for pharmaceutical and life science applications. The award-winning Echo 500 series liquid handlers and Portrait 630 reagent multi-spotters are used in nine of the 10 largest pharmaceutical companies, as well as in leading academic and research institutions and contract research organizations worldwide. The Labcyte acoustic droplet ejection technology has broad applications including compound management, assays, arraying, particle manufacturing, imaging mass spectrometry, and live-cell transfer. Labcyte also provides a range of unique microplate consumables. Labcyte has 29 issued U.S. patents, 3 issued European patents and additional international filings. For more information, visit www.labcyte.com

Posted under Collaborations, Equipment & Supplies, HT Screening, North America, Press Releases  |  Comments Off

Researchers from Novartis have published results using high-throughput screening (HTS) mass spectrometry (MS) techniques for the identification of enzyme inhibitors.

Last Updated on Tuesday, 23 October 2007 03:34 Written by Fred Tuesday, 23 October 2007 03:34

HTS techniques have become an important part of the drug discovery process allowing researchers to sift through massive libraries of compounds to find ‘hits’ that are active against a therapeutic target of interest.

This latest research, published as an early view article in the journal Analytical Chemistry by researchers from the Discovery Technologies and Diabetes and Metabolism divisions of Novartis‘ Institute for Biomedical Research, has assessed the usage of MS-HTS for large compound library screening.

Most functional activity-based programmes discover hits by detecting a change in the activity of a target enzyme in the presence of potential inhibitors or activators.

These changes in activity are commonly detected by using substrates that have been labelled with either radioisotopes or fluorescent probes.

According to the authors, labelled substrates do not always function exactly the same as native substrates and the developing methods to incorporate these labels can add significantly to method-development times.

MS can detect the products of enzymatic reactions without the need to label the substrates and the authors believe that this could potentially improve data quality achieved using the screens.

However, according to the authors, until recently throughput has been the major limiting factor in using MS for HTS and while using multiple instruments could be used to solve the problem, this would prove expensive.

“One contributing factor to the lack of large-scale [MS-HTS] studies is that instrumentation allowing unattended, fully automated analysis of thousands of samples per day with automated data analysis is not readily available,” write the authors.

The researchers looked at three different MS-HTS techniques, four-way parallel multiplexed electrospray liquid chromatography (LC) tandem MS (MUX-LC/MS/MS), four-way parallel staggered gradient liquid chromatography tandem MS (LC/MS/MS) and eight-way staggered flow injection MS/MS following 384-well plate solid phase extraction (SPE).

Each of these methods makes use of parallel autosamplers to enable more efficient use of the Waters Quattro Micro triple quadrupole MS instruments.

The techniques described in the paper allowed unattended analyses of screens of over 175,000 compounds, with a 384-well plate taking around 2 hours to analyse.

The researchers conducted two inhibitor screening campaigns using the techniques to show that they could be used reliably to detect inhibition of an (unnamed) enzyme by compounds in a library.

“In comparison to ultra-HTS methods (>100,000 wells per day), these MS methods are relatively slow (<10,000 wells per day), making single compound screening of more than 1m compounds impractical at this time,” write the authors.

However, because the screens require no custom labelling of reagents or the development of assays to measure labelling success considerable time can be saved during the method development phase.

When combined with ‘multiple compound per well’ screens that can increase analysis times by a factor of five, the MS screens compare favourably to traditional HTS methods.

“Although the MS analysis is relatively slow, overall timelines of MS-based screens from concept to hit list are similar to those of traditional HTS and ultra-HTS-based screens,” conclude the authors.

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Novartis weighs up label-free HTS for drug discovery

Last Updated on Tuesday, 2 October 2007 02:37 Written by Fred Tuesday, 2 October 2007 02:37

Researchers from Novartis have published results using high-throughput screening (HTS) mass spectrometry (MS) techniques for the identification of enzyme inhibitors.

HTS techniques have become an important part of the drug discovery process allowing researchers to sift through massive libraries of compounds to find ‘hits’ that are active against a therapeutic target of interest.

This latest research, published as an early view article in the journal Analytical Chemistry by researchers from the Discovery Technologies and Diabetes and Metabolism divisions of Novartis‘ Institute for Biomedical Research, has assessed the usage of MS-HTS for large compound library screening.

Most functional activity-based programmes discover hits by detecting a change in the activity of a target enzyme in the presence of potential inhibitors or activators.

These changes in activity are commonly detected by using substrates that have been labelled with either radioisotopes or fluorescent probes.

According to the authors, labelled substrates do not always function exactly the same as native substrates and the developing methods to incorporate these labels can add significantly to method-development times.

MS can detect the products of enzymatic reactions without the need to label the substrates and the authors believe that this could potentially improve data quality achieved using the screens.

However, according to the authors, until recently throughput has been the major limiting factor in using MS for HTS and while using multiple instruments could be used to solve the problem, this would prove expensive.

“One contributing factor to the lack of large-scale [MS-HTS] studies is that instrumentation allowing unattended, fully automated analysis of thousands of samples per day with automated data analysis is not readily available,” write the authors.

The researchers looked at three different MS-HTS techniques, four-way parallel multiplexed electrospray liquid chromatography (LC) tandem MS (MUX-LC/MS/MS), four-way parallel staggered gradient liquid chromatography tandem MS (LC/MS/MS) and eight-way staggered flow injection MS/MS following 384-well plate solid phase extraction (SPE).

Each of these methods makes use of parallel autosamplers to enable more efficient use of the Waters Quattro Micro triple quadrupole MS instruments.

The techniques described in the paper allowed unattended analyses of screens of over 175,000 compounds, with a 384-well plate taking around 2 hours to analyse.

The researchers conducted two inhibitor screening campaigns using the techniques to show that they could be used reliably to detect inhibition of an (unnamed) enzyme by compounds in a library.

“In comparison to ultra-HTS methods (>100,000 wells per day), these MS methods are relatively slow (<10,000 wells per day), making single compound screening of more than 1m compounds impractical at this time,” write the authors.

However, because the screens require no custom labelling of reagents or the development of assays to measure labelling success considerable time can be saved during the method development phase.

When combined with ‘multiple compound per well’ screens that can increase analysis times by a factor of five, the MS screens compare favourably to traditional HTS methods.

“Although the MS analysis is relatively slow, overall timelines of MS-based screens from concept to hit list are similar to those of traditional HTS and ultra-HTS-based screens,” conclude the authors. 

Posted under Discoveries, Innovations and Patents, Europe, HT Screening, North America, Press Releases  |  Comments Off

Cisbio has entered into a partnership agreement with Lumiphore to gain access to new fluorescence reporter molecules for use in its high throughput (HT) screening assays.

Last Updated on Tuesday, 11 September 2007 03:40 Written by Fred Tuesday, 11 September 2007 03:40

The agreement grants Cisbio exclusive access to the Lumi4 compounds for fluorescence applications in drug discovery, with Lumiphore free to develop partnerships to develop the technology in other areas.

Cisbio has played an important part in the development of HT drug screening technologies and has enabled researchers to conduct binding assays based on its homogenous time-resolved fluorescence (HTRF) system that combines Time-Resolved Fluorescence (TRF) and Fluorescence Resonance Energy Transfer (FRET).

“This agreement reinforces our position as real leaders the TR-FRET field, and is a very important step for us,” said François Degorce, head HTRF marketing and business development at Cisbio.

“We have plans to apply the technology in some of our current products as well as many of our mid-to-long term projects.”

Lumiphore’s lanthanide-based florescent reporter molecules will enable Cisbio to generate assays that provide more accurate results due to their long-emission lifetimes and strong luminescent properties that provide higher signal-to-noise (S/N) ratio.

“According the last figures I have, TR-FRET accounted for between 13 and 15 per cent of the HT screening market and the use of this technology is increasing rapidly compared with other competing technologies due to its increased sensitivity and robustness,” said Degorce.

During the TR-FRET process, fluorescent reporting molecules emit long-lived fluorescence signals when a fluorescence donor molecule binds to an acceptor / emitter molecule.

This is in contrast to many biological assay systems that use radioactive reagents, which pose health risks not just to the researchers themselves, but also to any live cell lines they may be conducting assays in.

The donor / acceptor molecules and proteins can be supplied from the company’s catalogue, or can be developed in-house or by Cisbio to enable researchers to study the binding of potential drug candidates.

This powerful new technology brings researchers increased assay sensitivity, higher throughput and greater reliability and flexibility than some other systems with Cisbio already providing a wide range of assays based on its original Europium cryptate technology.

These include assays for GPCRs (G Protein couple receptors), inflammation, metabolic diseases, central nervous system, biomarkers, kinase assays and other oncology related assays.

“The new technology will complement our existing catalogue offerings and quite a few of our existing assays will be upgraded to make use of it,” said Degorce.

In addition, the company also offer kits for monitoring bioprocesses such as protein or antibody engineering as well as enabling rapid quantification of antibodies during the production process.

“The Lumiphore technology is very bright and very stable, and will enable an upgraded version of the HTRF assays to be conducted on a expanded range of detection platforms,” said Degorce.

He continued by saying that because the Lumiphore technology uses a Terbium ion rather than the Europium ion that Cisbio currently uses a whole range of new possibilities could be accessed.

This could include the development of multicolour assays that use different donor-acceptor pairs that have very narrow excitation and emission bands.

The compounds resistance to bleaching allows samples to be archived allowing improved quality control comparisons between old and new data – a problem for more easily bleached organic dyes.

“When you run a screen with 100,000 samples and the detector system is not working properly, instead of throwing the plate away and having to start again because the lifetime of the dyes are too short lived to conduct the experiments the next day, you can store the plates in the fridge or freezer and study the plates the next week when the detector is back up and running,” said Degorce.

“Many luminescence screens actually destroy the reporter molecules such that any repeat read-outs are less sensitive than the first, with both our old Europium cryptate dye and this new Lumiphore technology you can just keep on reading the plates.”

Posted under Collaborations, Europe, HT Screening, North America, Press Releases  |  Comments Off

Cisbio international’s upcoming third annual “HTRF® In Drug Discovery” Symposium will be held September 26-28 in Sonoma, California

Last Updated on Thursday, 19 July 2007 02:13 Written by Fred Thursday, 19 July 2007 02:13

Over the course of three days, experts in drug screening from around the globe will assemble to discuss the latest applications of Cisbio’s proprietary HTRF® technology in the drug screening, target discovery and mechanistic fields.  A number of scientific sessions will specifically examine GPCR and kinase screening, cell-based biomarker detection and functional assays. 

 

HTRF® (homogenous time-resolved fluorescence) technology is a highly sensitive, robust technology for the detection of molecular interactions and is widely used for the high throughput stage of drug development. As part of its ongoing commitment to providing the highest quality technical support to HTRF® customers, Cisbio established its annual symposium in 2005 as a forum for exchanging ideas on drug screening and the latest in HTRF® innovation

Posted under Compound Libraries, Europe, HT Screening, Targeted Libraries  |  Comments Off

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