LabAutomation 2006 - SLAS
LabAutomation 2006 - SLAS
LabAutomation 2006 - SLAS
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TP19<br />
Mike Jones<br />
Cambridge Antibody Technology<br />
Cambridge, Cambridgeshire<br />
United Kingdom<br />
mike.jones@cambridgeantibody.com<br />
Where Laboratory Technologies Emerge and Merge<br />
Co-Author(s)<br />
Debbie Pattison<br />
Mark Lidament<br />
John Andrews<br />
Ruth Franks<br />
Stephen Clulow<br />
High Throughput 96 Well Purification of Biopharmaceuticals for Cell-Based Screening<br />
Large libraries of biopharmaceuticals, often containing over 1010 members, can be generated using molecular biology techniques.<br />
These biomolecules are expressed in prokaryotic and eukaryotic systems either as culture supernatants or in the periplasm of bacteria.<br />
By-products from mammalian and bacterial cells (eg. endotoxin) and reagents used to release proteins from the cells (eg. osmotic shock<br />
reagents) can have adverse effects on biochemical and cell-based assays. Using cell-based assays as primary screens improves the<br />
correlation between High Throughput Screening (HTS) results and secondary assays. It allows the identification of agonists at an early<br />
stage in the drug discovery cascade. We describe an automated high throughput 96 well purification process, using PhyNexus’ proprietary<br />
PhyTip column technology that provides ~2000 purified samples per week. This system has been applied to single-chain antibodies, IgGs<br />
and therapeutic proteins. By developing a high throughput 96 well sample purification process we can use cell-based assays for primary<br />
screening. We can also screen a larger number of samples in cell-based assays compared to the previous system, where throughput was<br />
limited by the capability to produce larger scale purified samples prior to testing in cell-assays. Purified samples are added to the cellbased<br />
assays and their activity is measured. The amount of inhibitor in these samples can be quantified using HTRF assays and the two<br />
results can be combined to enable samples to be ranked by potency.<br />
TP20<br />
Patricia Kasila<br />
PerkinElmer Life and Analytical Sciences<br />
Windham, New Hampshire<br />
patricia.kasila@perkinelmer.com<br />
Co-Author(s)<br />
Hao Xie<br />
Harry Harney<br />
Andy Raneri<br />
Greg Warner<br />
LANCETM cAMP + Evolution P3 + ViewLux = uHTS<br />
High Throughput Screening laboratories have been successfully automating assays to achieve higher throughput. To date only a small<br />
number of labs have been successfully able to go to even higher density formats, i.e., 1536-well microplates, due to the lack of precise<br />
liquid handling systems, homogeneous robust detection assays and high density detection equipment. Examples shown here will<br />
demonstrate that the LANCE cAMP assay, which allows for the direct measurement of receptor mediated adenylyl cyclase activation/<br />
inhibition in G-protein coupled receptors, the Evolution P3 liquid handling system and the ViewLux CCD Imager, will give comparable<br />
results with excellent precision for both 384-well and 1536-well formats.<br />
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