|Low Volume Liquid Handling of Organic Solvents for Compound Storage and Dissolution using mosquito|
Gillian Lewis, Tristan Cope, Joby Jenkins, David Gledhill & Rob Lewis
mosquito® is a positive displacement liquid handling system capable of pipetting solutions with a high degree of accuracy (to within 5%) and precision (CVs of <10%) in the nanolitre range. Here, we demonstrate mosquito’s ability to dispense accurately low volumes of organic solvents of low surface tension and to re-dissolve compounds which have been previously dried in storage wells.
|Improving the efficiency of 384 "mini-tube" technology using mosquito nanlolitre pipetting|
Joby Jenkins, Wayne Bowen, Rob Lewis & Chloe Milburn
384-well plate “mini-tube” consumables have been developed by automation companies such as REMP and The Automation Partnership (TAP) to hold compound samples divided into single-use storage tubes. Each tube holds 40 -75 µL, and is filled, sealed, stored and then thawed just once before use. However, when accessed by conventional pipetting technology, the tube’s practical working volume is as little as 20 µL, which means high compound wastage. The seal must also be pierced prior to pipetting to p
|Combinatorial synthesis of oxazolidin-2-ones using Fmoc-Ser(Trt)-OH, a library with high degree of diversity|
Marek Korinek, Vaclav Stastny and Miroslav Havranek
A library of oxazolidin-2-ones was prepared by solid phase synthesis. Crutial reaction steps were optimized. Highly divers building blocks were selected for the synthesis. All compounds were purified using preparative LC/MS with mass directed fraction collection.
|Stem Cell Derived Human Neurons in CNS Drug Discovery|
L Jackson, C Verastegui, C Wells, D Pau, A Nunn, A Finucane, M Lynch, S Boldt, L Gerrard, S.J. MacKenzie
Central nervous system disorders affect many people worldwide. To increase the quality and number of drugs available for treatments of CNS disorders such as Alzheimer’s, schizophrenia and anxiety, the success rate of CNS drug discovery programmes must be improved. The challenge in drug discovery is to develop more relevant assay systems to identify lead compounds for further development. Cell based screening assays currently used for CNS drug development involve primary cells or commercially
|Expression and Purification of PI3 Kinase alpha and Development of a Luminometric|
Brigitte Boldyreff1, Tine L. Rasmussen2, Hans H. Jensen2 and Olaf-Georg Issinger2
Human class Ia PI3 kinase p110alpha catalytic and p85alpha regulatory subunits were expressed and purified. A luminometric ATP depletion assay to measure PI3K activity was established and optimized. Furthermore, the autophosphorylation status of PI3K alpha was checked and the IC50 value for the known PI3K inhibitor wortmannin was determined. The assay is homogenous and is potentially suitable for high throughput screening to identify PI3K inhibitors.
|A genome-wide loss of function screen identifies new genes required for lung cancer cell proliferation|
PINNA G, ARAUJO N, MOROZOVA N, DETREZ MA & HAREL-BELLAN A
RNA interference (RNAi) provides an experimental tool for functional genomics analysis. We screened a genome-wide RNAi library to identify new genes involved in lung cancer cell proliferation. 72 % from the 257 genes identified were involved in general gene expression processes, 16 % were of unknown function or unrelated to proliferation, and 12% consisted of uncharacterized genes. These last sets of genes provide potential novel targets for lung cancer treatment.
|High Throughput Surface Tension Measurement|
Neil L. Campbell, Jon Weaver
Understanding the surface tension properties of aqueous surfactants and polymer materials is important for the pharmaceutical, coatings, paints, inks, surfactant and household products industries. The traditional methods of measuring surface tension are tedious and time-consuming, so only a few compounds can be measured by a single user per day.
|Lead Optimization Computational Protocol at PDB Scale to Rationally Optimize Attachments to a Given Kinase Inhibitor Scaffold|
Moriaud F., Henry T., Adcock S.A., Vorotynsev A.M., Martin L., Doppelt O.
We’ve used MED-SuMo to query and mine the Protein’s Surface Chemical Functions surrounding fragments of PDB ligands, seeking similarities with the kinase of interest (Vegfr DFGout, pdb code 2oh4, ligand code GIG) and collecting a library of 1129 unique fragments positioned in the vegfr’s active site and annotated with the counts of contacts and h-bonds. With them we optimize a substructure of the GIG ligand to find others DFGout ligands.
|Applications of EFC-based Protein-Protein Interaction Assays for Gi, Gs, and Gq-Coupled GPCR Screening|
Tom Wehrman, Kun Peng, Daniel Bassoni, Keith Olson
GPCR targets can be addresssed with a variety of technologies and they are increasingly being evaluated with multiple assay approaches. DiscoveRx has validated over 60 GPCR targets (Gi, Gs, Gq, Type A & Type B) in a novel generic cell-based assay format utilizing protein-protein interaction batween the GPCR C-terminus and b-arrestin2. Data outlining benefits provided for primary & secondary screening is presented.