LabAutomation 2006 - SLAS

LabAutomation2006
4:30 pm Tuesday, January 24, 2006 Track 3: High-Throughput Technologies Room: Learning Center
Wyndham Palm Springs Hotel
Maija Partanen
Co-Author(s)
Thermo Electron
Tori Richmond, Thermo Electron Corp., Biomarker Research
Helsinki, Finland
maija.partanen@thermo.com
Initiatives in Mass Spectrometry (BRIMS) Center
Merja Mehto, Arja Lamberg, Thermo Electron
M. Askenazi, Jennifer Sutton, Leo Bonilla, Thermo Electron Corp.,
Biomarker Research Initiatives in Mass Spectrometry (BRIMS) Center
Automated Profiling and Identification of Endogenous Peptidomic Markers in Human Plasma
Interest in biomarkers has experienced an explosion in recent years. A lot of attention is being paid to the clinical applications of looking
at the changes in MS patterns of the peptidome for detecting differences that may correlate to various disease states. The peptidome
of human plasma has been estimated to contain ~5000 unique peptides. A precise identification of the components of the peptidome is
critical not only to our understanding of the biology of disease states but also to our ability to discover robust markers for these states.
Here we describe a high throughput, automated method for profiling components of the plasma peptidome.
A robotic KingFisher 96 system was employed for transferring and step-wise manipulation of hydrophobic, surface-activated C-18
magnetic beads to selectively separate and enrich endogenous peptides and small proteins found in human plasma. To simulate a “time
course” experiment, plasma was spiked with increasing and decreasing amounts of known peptides at six different levels. The plate
containing the spiked and unspiked plasma was then processed on the Kingfisher 96 to capture and enrich endogenous and spiked
peptides. A portion of each processed sample was spotted onto a MALDI plate using a robotic processes station. Analysis was performed
on a vMALDI ion source coupled to an LTQ linear ion trap mass spectrometer.
A robust, high throughput and mass spec compatible method for enriching peptides from complex mixtures is demonstrated. This method
is free of albumin interference and reproducibly enriches endogenous peptides and small proteins.
9:00 am Wednesday, January 25, 2006 Track 3: High-Throughput Technologies Room: Learning Center
Wyndham Palm Springs Hotel
Paul Taylor
Boehringer Ingelheim
Ridgefield, Connecticutt
ptaylor2@rdg.boehringer-ingelheim.com
The Automation-Process Analysis Match: Case Studies in Optimizing Efficiencies
Over the last decade, the drug discovery community has experienced substantial changes in lead identification methods as increasing
levels of robotics have been incorporated. Clearly, not all automated solutions have provided optimal results. In many cases the reasons
are related not only to engineering aspects (which certainly have made considerable advances), but also to process or organizational
dependencies. In the competitive environment of high throughput screening, the challenge is finding an acceptable balance between
running campaigns rapidly and maintaining high quality data. Hence, automation tools have emerged for speeding up assay development,
rapidly accessing secure compound repositories, miniaturizing liquid handling steps, adapting to robotic systems, monitoring production
quality control and delivering robust dose response data. In every case, improvements have been the result of applying a combination of
automation, process analysis, statistics and a firm understanding of the underlying science. Real world highlights will be presented which
have resulted in improved optimization efficiencies and product delivery.
78