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2009 Meetings CalendarEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
(Source: Journal of Biomolecular Screening)
Source: Journal of Biomolecular Screening - November 12, 2009 Category: Molecular Biology Tags: Articles Source Type: journals

Product Focus: Analytical and Preparative InstrumentationEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
(Source: Journal of Biomolecular Screening)
Source: Journal of Biomolecular Screening - November 12, 2009 Category: Molecular Biology Tags: Articles Source Type: journals

Utilization of the TangoTM {beta}-Arrestin Recruitment Technology for Cell-Based EDG Receptor Assay Development and InterrogationEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Cellular assay development for the endothelial differentiation gene (EDG) family of G-protein-coupled receptors (GPCRs) and related lysophospholipid (LP) receptors is complicated by endogenous receptor expression and divergent receptor signaling. Endogenously expressed LP receptors exist in most tissue culture cell lines. these LP receptors, along with other endogenously expressed GPCRs, contribute to off-target signaling that can complicate interpretation of second-messenger-based cellular assay results. these receptors also activate a diverse and divergent set of cellular signaling pathways, necessitating the use of a va...
Source: Journal of Biomolecular Screening - November 12, 2009 Category: Molecular Biology Authors: Wetter, J. A., Revankar, C., Hanson, B. J. Tags: Articles Source Type: journals

A Continuous Protein Methyltransferase (G9a) Assay for Enzyme Activity Measurement and Inhibitor ScreeningEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The authors describe a continuous protein methylation assay using the G9a protein lysine methyltransferase and its substrate protein WIZ (widely interspaced zinc finger motifs). The assay is based on the coupling of the biotinylated substrate protein to streptavidin-coated FlashPlates and the transfer of radioactive methyl groups from the S-adenosyl-L-methionine to the substrate. The reaction progress is monitored continuously by proximity scintillation counting. The assay is very accurate, convenient, well suited for automation, and highly reproducible with standard errors in the range of 5%. Because of few pipetting step...
Source: Journal of Biomolecular Screening - November 12, 2009 Category: Molecular Biology Authors: Dhayalan, A., Dimitrova, E., Rathert, P., Jeltsch, A. Tags: Articles Source Type: journals

A Novel High-Throughput Screening Assay for HCN Channel Blocker Using Membrane Potential-Sensitive Dye and FLIPREmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this study, the authors report the development of a novel membrane potential-sensitive dye (MPSD) assay for HCN channel modulators that has been miniaturized into 384-well fluorescent imaging plate reader (FLIPR) high-throughput screening (HTS) format. When optimized (by cell plating density, plate type, cell recovery from cryopreservation), the wellto-well signal variability was low, with a Z' = 0.73 and coefficient of variation = 6.4%, whereas the MPSD fluorescence signal amplitude was -23,700 ± 1500 FLIPR3 relative fluorescence units (a linear relationship was found between HCN1 MPSD fluorescence signal and th...
Source: Journal of Biomolecular Screening - November 12, 2009 Category: Molecular Biology Authors: Vasilyev, D. V., Shan, Q. J., Lee, Y. T., Soloveva, V., Nawoschik, S. P., Kaftan, E. J., Dunlop, J., Mayer, S. C., Bowlby, M. R. Tags: Articles Source Type: journals

Virtual Screening Against {alpha}-CobratoxinEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
-Cobratoxin (Cbtx), the neurotoxin isolated from the venom of the Thai cobra Naja kaouthia , causes paralysis by preventing acetylcholine (ACh) binding to nicotinic acetylcholine receptors (nAChRs). In the current study, the region of the Cbtx molecule that is directly involved in binding to nAChRs is used as the target for anticobratoxin drug design. The crystal structure (1YI5) of Cbtx in complex with the acetylcholine binding protein (AChBP), a soluble homolog of the extracellular binding domain of nAChRs, was selected to prepare an -cobratoxin active binding site for docking. The amino acid residues (Ser182-Tyr192) of ...
Source: Journal of Biomolecular Screening - November 12, 2009 Category: Molecular Biology Authors: Utsintong, M., Talley, T. T., Taylor, P. W., Olson, A. J., Vajragupta, O. Tags: Articles Source Type: journals

Optimization of Assay Conditions fo r Dengue Virus Protease: Effect of Various Polyols and Nonionic DetergentsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The aim of this work was to perform a systematic study of the effect of nonionic detergents on the activity of the dengue virus NS2B-NS3 protease. To ensure a high activity of the protease, the assay procedures for the dengue virus and other flaviviral proteases published to date are performed in the presence of up to 35% glycerol, which does not represent the cellular physicochemical environment. In addition, the high viscosity of glycerol-containing solutions leads to various experimental problems in miniaturized assays. Using an internally quenched peptide substrate, the authors show that glycerol is not essential for e...
Source: Journal of Biomolecular Screening - November 12, 2009 Category: Molecular Biology Authors: Steuer, C., Heinonen, K. H., Kattner, L., Klein, C. D. Tags: Articles Source Type: journals

Inhibitors of RecA Activity Discovered by High-Throughput Screening: Cell-Permeable Small Molecules Attenuate the SOS Response in Escherichia coliEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The phenomenon of antibiotic resistance has created a need for the development of novel antibiotic classes with nonclassical cellular targets. Unfortunately, target-based drug discovery against proteins considered essential for in vitro bacterial viability has yielded few new therapeutic classes of antibiotics. Targeting the large proportion of genes considered nonessential that have yet to be explored by high-throughput screening, for example, RecA, can complement these efforts. Recent evidence suggests that RecA-controlled processes are responsible for tolerance to antibiotic chemotherapy and are involved in pathways tha...
Source: Journal of Biomolecular Screening - November 12, 2009 Category: Molecular Biology Authors: Wigle, T. J., Sexton, J. Z., Gromova, A. V., Hadimani, M. B., Hughes, M. A., Smith, G. R., Yeh, L.-A., Singleton, S. F. Tags: Articles Source Type: journals

CXCR2 Inverse Agonism Detected by Arrestin RedistributionEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
To study CXCR2 modulated arrestin redistribution, the authors employed arrestin as a fusion protein containing either the Aequorea victoria—derived enhanced green fluorescent protein (EGFP) or a recently developed mutant of eqFP611, a red fluorescent protein derived from Entacmaea quadricolor. This mutant, referred to as RFP611, had earlier been found to assume a dimeric quarternary structure. It was therefore employed in this work as a "tandem" (td) construct for pseudo monomeric fusion protein labeling. Both arrestin fusion proteins, containing either td RFP611 (Arr td RFP611) or enhanced green fluorescent protein ...
Source: Journal of Biomolecular Screening - November 12, 2009 Category: Molecular Biology Authors: Kredel, S., Wolff, M., Wiedenmann, J., Moepps, B., Nienhaus, G. U., Gierschik, P., Kistler, B., Heilker, R. Tags: Articles Source Type: journals

C5a-Stimulated Recruitment of {beta}-Arrestin2 to the Nonsignaling 7-Transmembrane Decoy Receptor C5L2Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
C5L2 (or GPR77) is a high-affinity receptor for the complement fragment C5a and its desarginated product, C5a-desArg. Unlike the classical C5a receptor CD88, C5L2 does not couple to intracellular G-protein-signaling pathways but is thought to function as a decoy receptor. The authors show that stimulation of C5L2 with C5a and C5a-desArg induces redistribution of green fluorescent protein—labeled β-arrestin2 to cytoplasmic vesicles. C3a and C3a-desArg were inactive in the β-arrestin translocation assay. Direct interaction of ligand-stimulated C5L2 with β-arrestin was confirmed using a novel β-galac...
Source: Journal of Biomolecular Screening - November 12, 2009 Category: Molecular Biology Authors: Van Lith, L. H.C., Oosterom, J., Van Elsas, A., Zaman, G. J.R. Tags: Articles Source Type: journals

Profiling Bioactivity of the ToxCast Chemical Library Using BioMAP Primary Human Cell SystemsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The complexity of human biology has made prediction of health effects as a consequence of exposure to environmental chemicals especially challenging. Complex cell systems, such as the Biologically Multiplexed Activity Profiling (BioMAP) primary, human, cell-based disease models, leverage cellular regulatory networks to detect and distinguish chemicals with a broad range of target mechanisms and biological processes relevant to human toxicity. Here the authors use the BioMAP human cell systems to characterize effects relevant to human tissue and inflammatory disease biology following exposure to the 320 environmental chemic...
Source: Journal of Biomolecular Screening - November 12, 2009 Category: Molecular Biology Authors: Houck, K. A., Dix, D. J., Judson, R. S., Kavlock, R. J., Yang, J., Berg, E. L. Tags: Articles Source Type: journals

Automated High-Content Screening for Compounds That Disassemble the Perinucleolar CompartmentEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
All solid malignancies share characteristic traits, including unlimited cellular proliferation, evasion of immune regulation, and the propensity to metastasize. The authors have previously described that a subnuclear structure, the perinucleolar compartment (PNC), is associated with the metastatic phenotype in solid tumor cancer cells. The percentage of cancer cells that contain PNCs (PNC prevalence) is indicative of the malignancy of a tumor both in vitro and in vivo, and thus PNC prevalence is a marker that reflects metastatic capability in a population of tumor cells. Although the function of the PNC remains to be deter...
Source: Journal of Biomolecular Screening - November 12, 2009 Category: Molecular Biology Authors: Norton, J. T., Titus, S. A., Dexter, D., Austin, C. P., Zheng, W., Huang, S. Tags: Articles Source Type: journals

In Memoriam: Tony J. Beugelsdijk (1949--2009)Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
(Source: Journal of Biomolecular Screening)
Source: Journal of Biomolecular Screening - November 12, 2009 Category: Molecular Biology Tags: Articles Source Type: journals

2009-2010 Meetings CalendarEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
(Source: Journal of Biomolecular Screening)
Source: Journal of Biomolecular Screening - October 1, 2009 Category: Molecular Biology Tags: Articles Source Type: journals

Product Focus: Software, Databases, and Information ServicesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
(Source: Journal of Biomolecular Screening)
Source: Journal of Biomolecular Screening - October 1, 2009 Category: Molecular Biology Tags: Articles Source Type: journals

Establishing Quality Assurance Criteria for Serial Dilution Operations on Liquid-Handling EquipmentEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Since the advent of high-throughput screening (HTS) in the early 1990s, parallel multichannel liquid handlers have become a mainstay in every drug discovery setting. Although several peer-reviewed publications have discussed methods and criteria for stamping multiwell copies, there is very little information about establishing a standard operating procedure (SOP) for standard (microliter-level) serial dilutions of compounds used in dose-response experiments. The authors discuss the 4 main criteria any serial dilution process must pass (accuracy, precision, fold dilution, and outliers) and the process for establishing thres...
Source: Journal of Biomolecular Screening - October 1, 2009 Category: Molecular Biology Authors: Popa-Burke, I., Lupotsky, B., Boyer, J., Gannon, W., Hughes, R., Kadwill, P., Lyerly, D., Nichols, J., Nixon, E., Rimmer, D., Saiz-Nicolas, I., Sanfiz-Pinto, B., Holland, S. Tags: Articles Source Type: journals

Correction for Interference by Test Samples in High-Throughput AssaysEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In high-throughput biochemical assays performed in multiwell plates, the effect of test samples on the activity of the biochemical system is usually measured by optical means such as absorbance, fluorescence, luminescence, or scintillation counting. The test sample often causes detection interference when it remains in the well during the measurement. Interference may be due to light absorption, fluorescence quenching, sample fluorescence, chemical interaction of the sample with a detection reagent, or depression of the meniscus. A simple method is described that corrects for such interference well by well. The interferenc...
Source: Journal of Biomolecular Screening - October 1, 2009 Category: Molecular Biology Authors: Shapiro, A. B., Walkup, G. K., Keating, T. A. Tags: Articles Source Type: journals

A Primer on Screening Data ManagementEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
A drug discovery startup company or academic lab entering the screening arena faces numerous challenges as it tries to manage the large quantity of data generated by a typical drug discovery screening campaign. Although there are sophisticated off-the-shelf software solutions available, their use requires substantial forethought and attention to detail if the data they capture are to be of sufficient quality to serve the various purposes to which it will be put. For newcomers to the field of screening data management in particular, the problem is compounded by a lack of literature covering the practical aspects of managing...
Source: Journal of Biomolecular Screening - October 1, 2009 Category: Molecular Biology Authors: Palmer, M., Kremer, A., Terstappen, G. C. Tags: Articles Source Type: journals

Neutralizing Antibodies of Botulinum Neurotoxin Serotype A Screened from a Fully Synthetic Human Antibody Phage Display LibraryEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The botulinum neurotoxins (BoNTs) produced by Clostridium botulinum are the most poisonous protein substances known. The neutralizing antibodies against botulinum neurotoxin can effectively prevent and cure the toxicosis. Using purified Hc fragments of botulinum neurotoxin serotype A (BoNT/A-Hc) as antigen, 2 specific neutralizing antibodies mapping different epitopes were selected from a fully synthetic human antibody library. The 2 antibodies can effectively inhibit the binding between BoNT/A-Hc and differentiated PC-12 cells in vitro, and the neutralization was evaluated in vivo. Although no single mAb completely protec...
Source: Journal of Biomolecular Screening - October 1, 2009 Category: Molecular Biology Authors: Yu, R., Wang, S., Yu, Y.-z., Du, W.-s., Yang, F., Yu, W.-y., Sun, Z.-w. Tags: Articles Source Type: journals

Test System for Trifunctional Antibodies in 3D MCTS CultureEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The aim of the present study was to assess the feasibility of a 3D tumor cell culture model, that is, multicellular tumor spheroids (MCTSs) as an adequate model for micrometastases and therefore as a pharmacological model for efficacy testing of trifunctional therapeutic antibodies. Unlike conventional monolayer cultures, spheroids allow researchers to study parameters, such as 3D cell shape, 3D cell arrangement and microenvironment, and penetration efficiency of defense cells that may largely influence the efficacy of antibody treatment in vivo. The authors established a long-term coculture of human MCTSs with peripheral ...
Source: Journal of Biomolecular Screening - October 1, 2009 Category: Molecular Biology Authors: Hirschhaeuser, F., Leidig, T., Rodday, B., Lindemann, C., Mueller-Klieser, W. Tags: Articles Source Type: journals

Novel In Vitro Protein Fragment Complementation Assay Applicable to High-Throughput Screening in a 1536-Well FormatEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Protein-protein interactions (PPIs) play key roles in all cellular processes and hence are useful as potential targets for new drug development. To facilitate the screening of PPI inhibitors as anticancer drugs, the authors have developed a high-throughput screening (HTS) system using an in vitro protein fragment complementation assay (PCA) with monomeric Kusabira-Green fluorescent protein (mKG). The in vitro PCA system was established by the topological formation of a functional complex between 2 split inactive mKG fragments fused to target proteins, which fluoresces when 2 target proteins interact to allow complementatio...
Source: Journal of Biomolecular Screening - October 1, 2009 Category: Molecular Biology Authors: Hashimoto, J., Watanabe, T., Seki, T., Karasawa, S., Izumikawa, M., Seki, T., Iemura, S.-I., Natsume, T., Nomura, N., Goshima, N., Miyawaki, A., Takagi, M., Shin-Ya, K. Tags: Articles Source Type: journals

Live-Cell Imaging of Caspase Activation for High-Content ScreeningEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Caspases are central to the execution of programmed cell death, and their activation constitutes the biochemical hallmark of apoptosis. In this article, the authors report the successful adaptation of a high-content assay method using the DEVDNucView488TM fluorogenic substrate, and for the first time, they show caspase activation in live cells induced by either drugs or siRNA. The fluorogenic substrate was found to be nontoxic over an exposure period of several days, during which the authors demonstrate automated imaging and quantification of caspase activation of the same cell population as a function of time. Overexpress...
Source: Journal of Biomolecular Screening - October 1, 2009 Category: Molecular Biology Authors: Antczak, C., Takagi, T., Ramirez, C. N., Radu, C., Djaballah, H. Tags: Articles Source Type: journals

DetecTiff(C): A Novel Image Analysis Routine for High-Content Screening MicroscopyEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this article, the authors describe the image analysis software DetecTiff©, which allows fully automated object recognition and quantification from digital images. The core module of the LabView©-based routine is an algorithm for structure recognition that employs intensity thresholding and size-dependent particle filtering from microscopic images in an iterative manner. Detected structures are converted into templates, which are used for quantitative image analysis. DetecTiff © enables processing of multiple detection channels and provides functions for template organization and fast interpretation of acq...
Source: Journal of Biomolecular Screening - October 1, 2009 Category: Molecular Biology Authors: Gilbert, D. F., Meinhof, T., Pepperkok, R., Runz, H. Tags: Articles Source Type: journals

A New Simple Cell-Based Homogeneous Time-Resolved Fluorescence QRET Technique for Receptor-Ligand Interaction ScreeningEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this article, a single-label separation-free fluorescence technique is presented as a potential screening method for cell-based receptor antagonists and agonists.The time-resolved fluorescence technique, quenching resonance energy transfer (QRET), relies on a single-labeled binding partner in combination with a soluble quencher. The quencher efficiently suppresses the luminescence of the unbound labeled ligand, whereas the luminescence of the bound fraction is not affected. This approach allows the development of cell-based screening assays in a simple and cost-effective manner. The authors have applied the technique to...
Source: Journal of Biomolecular Screening - October 1, 2009 Category: Molecular Biology Authors: Harma, H., Rozwandowicz-Jansen, A., Martikkala, E., Frang, H., Hemmila, I., Sahlberg, N., Fey, V., Perala, M., Hanninen, P. Tags: Articles Source Type: journals

Development and Applications of a Broad-Coverage, TR-FRET-Based Kinase Binding Assay PlatformEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The expansion of kinase assay technologies over the past decade has mirrored the growing interest in kinases as drug targets. As a result, there is no shortage of convenient, fluorescence-based methods available to assay targets that span the kinome. The authors recently reported on the development of a non-activity-based assay to characterize kinase inhibitors that depended on displacement of an Alexa Fluor® 647 conjugate of staurosporine (a "tracer") from a particular kinase. Kinase inhibitors were characterized by a change in fluorescence lifetime of the tracer when it was bound to a kinase relative to when it was d...
Source: Journal of Biomolecular Screening - October 1, 2009 Category: Molecular Biology Authors: Lebakken, C. S., Riddle, S. M., Singh, U., Frazee, W. J., Eliason, H. C., Gao, Y., Reichling, L. J., Marks, B. D., Vogel, K. W. Tags: Articles Source Type: journals

Mechanism-Based Inhibition: Deriving KI and kinact Directly from Time-Dependent IC50 ValuesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The potential of enzyme inhibition of a drug is frequently quantified in terms of IC50 values. Although this is a suitable quantity for reversible inhibitors, concerns arise when dealing with irreversible or mechanism-based inhibitors (MBIs). IC50 values of MBIs are time dependent, causing serious problems when aiming at ranking different compounds with respect to their inhibitory potential. As a consequence, most studies and ranking schemes related to MBIs rely on the inhibition constant (KI) and the rate of enzyme inactivation (kinact) rather than on IC50 values. In this article, the authors derive a novel relation betwe...
Source: Journal of Biomolecular Screening - October 1, 2009 Category: Molecular Biology Authors: Krippendorff, B.-F., Neuhaus, R., Lienau, P., Reichel, A., Huisinga, W. Tags: Articles Source Type: journals

Society UpdatesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
(Source: Journal of Biomolecular Screening)
Source: Journal of Biomolecular Screening - October 1, 2009 Category: Molecular Biology Tags: Articles Source Type: journals

Oral Abstracts from the Society of Biomolecular Sciences 15th Annual Conference and ExhibitionEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
(Source: Journal of Biomolecular Screening)
Source: Journal of Biomolecular Screening - August 31, 2009 Category: Molecular Biology Tags: Article Source Type: journals

2009 Meetings CalendarEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
(Source: Journal of Biomolecular Screening)
Source: Journal of Biomolecular Screening - August 31, 2009 Category: Molecular Biology Tags: Article Source Type: journals

Product Focus: Microplates, Assay Reagents, Screening Consumables, and KitsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
(Source: Journal of Biomolecular Screening)
Source: Journal of Biomolecular Screening - August 31, 2009 Category: Molecular Biology Tags: Article Source Type: journals

A High-Throughput Method to Identify Novel Senescence-Inducing CompoundsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Cellular senescence is a persistently growth-arrested phenotype in normal and transformed cells induced by noncytotoxic stress. Cytostasis as a method of cancer treatment has recently generated significant interest. Research into the induction of cellular senescence as cancer therapy has been hindered by a lack of compounds that efficiently induce this response. The authors describe a semiautomated high-throughput method to identify library compounds that induce senescence using prostate cancer cells cultured in 96-well plates. Primary hits are identified by low cell numbers after 3 days in culture, measured by Hoechst 333...
Source: Journal of Biomolecular Screening - August 31, 2009 Category: Molecular Biology Authors: Ewald, J. A., Peters, N., Desotelle, J. A., Hoffmann, F. M., Jarrard, D. F. Tags: Article Source Type: journals

Multiplexing of Pathway-Specific {beta}-Lactamase Reporter Gene Assays by Optical Coding With Qtracker(R) NanocrystalsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Reporter assays are widely used in research and drug discovery for analysis of signaling pathways in a cell-based format. Traditionally, reporter gene assays are run in a single-parameter mode, interrogating only 1 pathway per sample. To enable more complex assay formats for pathway analysis, the authors developed a multiplexed reporter cell-based assay that combines optical encoding with a β-lactamase reporter gene readout. The optical encoding is achieved by peptide-mediated delivery of quantum dots into reporter cell lines. Using different quantum dots, the authors were able to simultaneously analyze multiple signa...
Source: Journal of Biomolecular Screening - August 31, 2009 Category: Molecular Biology Authors: Machleidt, T., Whitney, P., Bi, K. Tags: Article Source Type: journals

High-Throughput, Cell-Free, Liposome-Based Approach for Assessing In Vitro Activity of Lipid KinasesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Lipid kinases are central players in lipid signaling pathways involved in inflammation, tumorigenesis, and metabolic syndrome. A number of these kinase targets have proven difficult to investigate in higher throughput cell-free assay systems. This challenge is partially due to specific substrate interaction requirements for several of the lipid kinase family members and the resulting incompatibility of these substrates with most established, homogeneous assay formats. Traditional, cell-free in vitro investigational methods for members of the lipid kinase family typically involve substrate incorporation of [-32P] and resolu...
Source: Journal of Biomolecular Screening - August 31, 2009 Category: Molecular Biology Authors: Demian, D. J., Clugston, S. L., Foster, M. M., Rameh, L., Sarkes, D., Townson, S. A., Yang, L., Zhang, M., Charlton, M. E. Tags: Article Source Type: journals

Identification and Characterization of Novel Tissue-Nonspecific Alkaline Phosphatase Inhibitors with Diverse Modes of ActionEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
This article provides an example of a success where the Roadmap Initiative collaborative model, sponsored by the National Institutes of Health, brought together a deep knowledge of target biology from a principal investigator’s laboratory, a well-designed compound collection from the MLSMR, and an industrial-level screening facility and staff at the MLSCN center to identify pharmacologically active compounds, with outstanding selectivity data from a panel of more than 200 publicly accessible assays, through a high-throughput screen. (Journal of Biomolecular Screening 2009:824-837) (Source: Journal of Biomolecular Screening)
Source: Journal of Biomolecular Screening - August 31, 2009 Category: Molecular Biology Authors: Sergienko, E., Su, Y., Chan, X., Brown, B., Hurder, A., Narisawa, S., Millan, J. L. Tags: Article Source Type: journals

Pharmacological Characterization of Receptor Redistribution and {beta}-Arrestin Recruitment Assays for the Cannabinoid Receptor 1Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Receptor redistribution and β-arrestin recruitment assays provide a G-protein-subtype-independent method to measure ligand-stimulated activation of G-protein-coupled receptors. In particular β-arrestin assays are becoming an increasingly popular tool for drug discovery. The authors have compared a high-content-imaging-based Redistribution® assay and 2 nonimaging-based β-arrestin recruitment assays, TangoTM and PathHunter TM, for the cannabinoid receptor 1. Inasmuch as all 3 assays use receptors that are modified at the C-terminus, the authors verified their pharmacology via detection of Gi coupling of th...
Source: Journal of Biomolecular Screening - August 31, 2009 Category: Molecular Biology Authors: van der Lee, M. M.C., Blomenrohr, M., van der Doelen, A. A., Wat, J. W.Y., Smits, N., Hanson, B. J., van Koppen, C. J., Zaman, G. J.R. Tags: Article Source Type: journals

A Homogeneous Fluorescent Live-Cell Assay for Measuring 7-Transmembrane Receptor Activity and Agonist Functional Selectivity Through Beta-Arrestin RecruitmentEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Seven-transmembrane (7TM) receptors play an essential role in the regulation of a wide variety of physiological processes, making them one of the top target classes for pharmaceuticals. 7TM receptor function is mediated and modulated through 2 primary processes: G-protein and beta-arrestin signaling. Classically, it has been recognized that these 2 processes can interact with one another during 7TM receptor desensitization, but it has more recently been recognized that these 2 processes can also act independently of one another and can activate parallel signaling pathways. As such, the methods used to interrogate 7TM recep...
Source: Journal of Biomolecular Screening - August 31, 2009 Category: Molecular Biology Authors: Hanson, B. J., Wetter, J., Bercher, M. R., Kopp, L., Fuerstenau-Sharp, M., Vedvik, K. L., Zielinski, T., Doucette, C., Whitney, P. J., Revankar, C. Tags: Article Source Type: journals

Identification of Surrogate Agonists and Antagonists for Orphan G-Protein-Coupled Receptor GPR139Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
GPR139 is an orphan G-protein-coupled receptor (GPCR) that is expressed nearly exclusively in the central nervous system and may play a role in the control of locomotor activity. The signal transduction pathway and pharmacological function of GPR139, however, are still controversial due to the lack of natural or synthetic ligands. The authors report the characterization of human GPR139 signaling pathway and identification of surrogate agonists and antagonists. In both transient and stable transfections of HEK293F cells, overexpression of GPR139 increased basal intracellular cAMP concentrations compared to control cells. Fu...
Source: Journal of Biomolecular Screening - August 31, 2009 Category: Molecular Biology Authors: Hu, L. A., Tang, P. M., Eslahi, N. K., Zhou, T., Barbosa, J., Liu, Q. Tags: Article Source Type: journals

Development of Generic Calcium Imaging Assay for Monitoring Gi-Coupled Receptors and G-Protein InteractionEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
G-protein-coupled receptors (GPCRs) are important therapeutic targets for many areas of drug research and development. Although chimeric G16 proteins are valuable tools for detecting the activation of Gi/o-coupled receptors, the details of the activation process remain unclear. The authors introduce a series of chimeras that combine both G16 and G i/o (G16/o, G16/i2, and G 16/i3) into a well-established transient expression system to examine the ability of these chimeras to interact with D2 long-form (D 2L) dopamine and 5-HT1A serotonin receptors. The pEC 50 data obtained for known agonists were similar to results from pre...
Source: Journal of Biomolecular Screening - August 31, 2009 Category: Molecular Biology Authors: Ueda, T., Ugawa, S., Ishida, Y., Hondoh, A., Shimada, S. Tags: Article Source Type: journals

Population Patch-Clamp Electrophysiology Analysis of Recombinant GABAA {alpha}1{beta}3{gamma}2 Channels Expressed in HEK-293 CellsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
-Amino butyric acid (GABA)—activated Cl— channels are critical mediators of inhibitory postsynaptic potentials in the CNS. To date, rational design efforts to identify potent and selective GABAA subtype ligands have been hampered by the absence of suitable high-throughput screening approaches. The authors describe 384-well population patch-clamp (PPC) planar array electrophysiology methods for the study of GABAA receptor pharmacology. In HEK293 cells stably expressing human 1β32 GABAA channels, GABA evoked outward currents at 0 mV of 1.05 ± 0.08 nA, measured 8 s post GABA addition. The IGABA was lin...
Source: Journal of Biomolecular Screening - August 31, 2009 Category: Molecular Biology Authors: Hollands, E. C., Dale, T. J., Baxter, A. W., Meadows, H. J., Powell, A. J., Clare, J. J., Trezise, D. J. Tags: Article Source Type: journals

Stem Cells in Drug Discovery, Tissue Engineering, and Regenerative Medicine: Emerging Opportunities and ChallengesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Stem cells, irrespective of their origin, have emerged as valuable reagents or tools in human health in the past 2 decades. Initially, a research tool to study fundamental aspects of developmental biology is now the central focus of generating transgenic animals, drug discovery, and regenerative medicine to address degenerative diseases of multiple organ systems. This is because stem cells are pluripotent or multipotent cells that can recapitulate developmental paths to repair damaged tissues. However, it is becoming clear that stem cell therapy alone may not be adequate to reverse tissue and organ damage in degenerative d...
Source: Journal of Biomolecular Screening - August 31, 2009 Category: Molecular Biology Authors: Nirmalanandhan, V. S., Sittampalam, G. S. Tags: Article Source Type: journals

Society UpdatesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
(Source: Journal of Biomolecular Screening)
Source: Journal of Biomolecular Screening - August 31, 2009 Category: Molecular Biology Tags: Article Source Type: journals

2009 Meetings CalendarEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
(Source: Journal of Biomolecular Screening)
Source: Journal of Biomolecular Screening - July 16, 2009 Category: Molecular Biology Tags: Article Source Type: journals

Product Focus: Screening Robotics and AutomationEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
(Source: Journal of Biomolecular Screening)
Source: Journal of Biomolecular Screening - July 16, 2009 Category: Molecular Biology Tags: Article Source Type: journals

Primary Leukocyte Screens for Innate Immune AgonistsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The innate immune system of mammals is a key defense mechanism against invading foreign pathogens. Innate immune stimulants may have applications as vaccine adjuvants as well as in the treatment of cancer and some viral diseases, and clinical studies have been performed using agonists of Toll-like receptors (TLRs) 7, 8, and 9. The high-throughput screens for such agonists have typically relied on the overexpression of a single TLR gene in an immortalized cell line and are inherently artificial systems that are restricted to the identification of agonists for a single receptor. The authors describe 2 assays for the identifi...
Source: Journal of Biomolecular Screening - July 16, 2009 Category: Molecular Biology Authors: Goodchild, A., Nopper, N., Craddock, A., Law, T., King, A., Fanning, G., Rivory, L., Passioura, T. Tags: Article Source Type: journals

Use of Reduced Temperature Cell Pausing to Enhance Flexibility of Cell-Based AssaysEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Construction and supply of cell-based reagents for in vitro plate-based screens are often highlighted as a bottleneck within drug discovery. Recent years have seen the successful application of both cryopreservation and automation to increase the capacity and flexibility of cell provision. However, routine cell culture remains a fixed experimental process that requires cells to be prepared and used at specific times. We have investigated the potential of reduced temperature incubation to be used as a simple methodology for stopping and starting cell growth and introduce further flexibility into cell provision. Our results ...
Source: Journal of Biomolecular Screening - July 16, 2009 Category: Molecular Biology Authors: Wise, H., Abel, P. W., Cawkill, D. Tags: Article Source Type: journals

Overcoming Problems of Compound Storage in DMSO: Solvent and Process AlternativesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The common practice of preparing storage libraries of compounds in 100% DMSO solution well in advance of bioassay brings with it difficulties that affect the accuracy of the data obtained. This publication presents a series of studies done on a subset of compounds that are difficult to bioassay because they precipitate from DMSO solution. These compounds are members of a frequently used, diverse compound library of the sort commonly used in the high-throughput screening (HTS) environment. Experiments were performed to determine the concentration of drug in solution above the precipitate, observe the time course and effect ...
Source: Journal of Biomolecular Screening - July 16, 2009 Category: Molecular Biology Authors: Waybright, T. J., Britt, J. R., McCloud, T. G. Tags: Article Source Type: journals

Buffer Optimization of Thermal Melt Assays of Plasmodium Proteins for Detection of Small-Molecule LigandsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In the past decade, thermal melt/thermal shift assays have become a common tool for identifying ligands and other factors that stabilize specific proteins. Increased stability is indicated by an increase in the protein's melting temperature (Tm). In optimizing the assays for subsequent screening of compound libraries, it is important to minimize the variability of Tm measurements so as to maximize the assay's ability to detect potential ligands. The authors present an investigation of Tm variability in recombinant proteins from Plasmodium parasites. Ligands of Plasmodium proteins are particularly interesting as potential s...
Source: Journal of Biomolecular Screening - July 16, 2009 Category: Molecular Biology Authors: Crowther, G. J., Napuli, A. J., Thomas, A. P., Chung, D. J., Kovzun, K. V., Leibly, D. J., Castaneda, L. J., Bhandari, J., Damman, C. J., Hui, R., Hol, W. G.J., Buckner, F. S., Verlinde, C. L.M.J., Zhang, Z., Fan, E., van Voorhis, W. C. Tags: Article Source Type: journals

Plate-Based Diversity Selection Based on Empirical HTS Data to Enhance the Number of Hits and Their Chemical DiversityEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Typically, screening collections of pharmaceutical companies contain more than a million compounds today. However, for certain high-throughput screening (HTS) campaigns, constraints posed by the assay throughput and/or the reagent costs make it impractical to screen the entire deck. Therefore, it is desirable to effectively screen subsets of the collection based on a hypothesis or a diversity selection. How to select compound subsets is a subject of ongoing debate. The authors present an approach based on extended connectivity fingerprints to carry out diversity selection on a per plate basis (instead of a per compound bas...
Source: Journal of Biomolecular Screening - July 16, 2009 Category: Molecular Biology Authors: Sukuru, S. C. K., Jenkins, J. L., Beckwith, R. E.J., Scheiber, J., Bender, A., Mikhailov, D., Davies, J. W., Glick, M. Tags: Article Source Type: journals

Efficient Elimination of Nonstoichiometric Enzyme Inhibitors from HTS Hit ListsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this study, the effect of such unwanted inhibitors on several different enzyme targets was assessed based on screening results for over a million compounds. In particular, the shift in potency on variation of enzyme concentration was used as a means to identify nonstoichiometric inhibitors among the screening hits. These potency shifts depended on both compound structure and target enzyme. The approach was confirmed by statistical analysis of thousands of dose-response curves, which showed that the potency of competitive and therefore clearly stoichiometric inhibitors was not affected by increasing enzyme concentration....
Source: Journal of Biomolecular Screening - July 16, 2009 Category: Molecular Biology Authors: Habig, M., Blechschmidt, A., Dressler, S., Hess, B., Patel, V., Billich, A., Ostermeier, C., Beer, D., Klumpp, M. Tags: Article Source Type: journals

Development of a Fluorescence-Based, Ultra High-Throughput Screening Platform for Nanoliter-Scale Cytochrome P450 MicroarraysEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Cytochrome P450 enzyme (CYP450s) assays are critical enzymes in early-stage lead discovery and optimization in drug development. Currently available fluorescence-based reaction assays provide a rapid and reliable method for monitoring CYP450 enzyme activity but are confined to medium-throughput well-plate systems. The authors present a high-throughput, integrated screening platform for CYP450 assays combining enzyme encapsulation techniques, microarraying methods, and wide-field imaging. Alginate-containing microarrays consisting of up to 1134 CYP450 reaction elements were fabricated on functionalized glass slides (reactio...
Source: Journal of Biomolecular Screening - July 16, 2009 Category: Molecular Biology Authors: Sukumaran, S. M., Potsaid, B., Lee, M.-Y., Clark, D. S., Dordick, J. S. Tags: Article Source Type: journals