|TDS LIMS: A Platform for Comprehensive Management and Analysis of Screening Data|
Karol Kozak, Marta Kozak, Jan Wagner, Hannes Grabner, Kerstin Korn, Eugenio Fava, Marit Biesold, Claudia Moebius, Anett Lohman and Ebrhard Krausz
Here we present a web-based bioinformatics solution for the management and analysis of all areas of cell-based screen experimentation: the Laboratory Information Management System of the Technology Development Studio (TDS-LIMS). TDS-LIMS is a software package consisting of the Library Checker, Library Browser, Scheduling Kit, Screen Browser and Screen Publisher.
|A High Content Primary Screening Assay Determining ERK and JNK Activation|
Paul Wylie and Wayne Bowen
Here we present a rapid method for determining ERK and JNK activation, demonstrated using time course, concentration-dependence data and the effect of the MEK1/2 inhibitor UO126 on ERK activation obtained using the Acumen Explorer™ laser scanning cytometer analysing 384 well plates.
|Cell Cycle Analysis Using Microplate Cytometry: A Comparison of Laser and Dye Combinations|
Tristan Cope, Christopher Lupton, Paul Wylie, Jeff Hung and Wayne Bowen
For improved screening capability, we have developed a cell cycle analysis method using an Acumen Explorer fluorescence microplate cytometer, capable of reading an entire 384 well microplate in about 10 minutes. The method can perform such analyses on fixed cells in situ, markedly simplifying sample preparation.
|Determination of Cell Colony Formation in a High Content Screening Assay|
David Pole and Wayne Bowen
Here, laser scanning microplate cytometry has been used to provide an automated high content readout of the effects of cytostatic agents on colony formation. This approach determines colony number through the application of a spherical volume algorithm. This permits the differentiation of cytostatic effects where the number of colonies and size remains constant and cytotoxic effects where the size and number may be reduced.
|Automated Nanolitre Hit Automated Nanolitre Hit - Picking using a Mosquito® X1 Low Volume Pipettor|
Joby Jenkins, Rob Lewis, Tristan Cope, Wayne Bowen
The mosquito® X1 offers precision sampling of any individual well in 48-, 96-, 384- or 1536-well plates. This enables researchers to quickly select small volumes of “hits” from primary screening plates and transfer them directly to the next screening stage without further dilution. mosquito® X1's disposable pipette tips guarantee zero cross-contamination.
|The use of Benzyl Esters in the Synthesis of 1ß-O-Acyl Glucuronides by Selective Acylation|
Elizabeth R. Bowkett, B. Kevin Park and Andrew V. Stachulski
Acyl glucuronides are key phase 2 metabolites for many carboxylic acid-containing drugs, notably of non-steroidal anti-inflammatory agents (NSAIDs).1 It is important to synthesise acyl glucuronides in pure form, and especially as single 1ß-anomers, for bioevaluation during drug discovery and development. Acyl glucuronides are reactive species: they may react by hydrolysis or displacement with other nucleophiles or by acyl migration followed by condensation with amines.
|Multicolor Detection of CD45 on Human Monocytes with Quantum Dots in a High-Throughput Format|
Paul Wylie, John Budd and Marcel Bruchez
Multicolor quantum dot probes emitting from 525 to 705 nm can be effectively excited with a single 488 nm laser, resulting in signals equal to the best organic dyes, but that are cleanly discriminated with simple color filters. Using a plate based 4-color laser scanning system with a single 488 or 405 nm laser for excitation, we have detected the binding of quantum dot conjugates to CD45 on human monocytes.
|Optimizing the hERG Assay on the PatchXpress 7000A System for Compounds that Demonstrate Non-specific Binding|
Iris Yang, Naibo Yang, Cathy Smith-Maxwell, Claire Quinn, Kirsty Macfarlane and David Yamane
Eight compounds, astemizole, pimozide, dofetilide, cisapride, terfenadine, flunarizine, quinidine and imipramine were used to evaluate the effect of different assay conditions to overcome the effects of non-specific binding in our hERG assay on the PatchXpress 7000A system.
|Reducing the Synthetic Burdens of Lead Structure Optimization: A Novel Software-Aided Approach|
Karim Kassam, Ryan Sasaki and Michel R.J. Hachey
At the later stages of drug optimization it is beneficial to augment the select ADME properties of leads by making small structural changes that do not introduce any negative effects on the activity or toxicity profiles. By combining physicochemical property predictors and a critically evaluated database of biologically-acceptable substituents, the synthetic chemist can evaluate and reduce the number of derivative compounds that need to be synthesized to achieve optimal drug-like properties.