omation mbers - Society for Laboratory Automation and Screening

8:30 am Thursday, February 5 Microchip – Separations Room A4
Don L. DeVoe
Calibrant BioSystems, Inc.
7507 Standish Place
Rockville, Maryland 20855
ddev@calibrant.com
2D PAGE on a Chip: Multidimensional Protein Separations via High Throughput Microfluidics
An automated system based on a microfluidic analog of traditional 2D PAGE has been developed for ultra-high
throughput protein analysis. For the analysis of complex protein mixtures, two-dimensional polyacrylamide gel
electrophoresis (2D PAGE) remains the method of choice for separating more than thousands of proteins prior
to mass spectrometry (MS), with proteins separated by charge and size in a 2D gel. Despite the selectivity and
sensitivity of 2D PAGE, this technique as practiced today is the collection of manually intensive procedures.
Casting of gels, application of samples, running of gels, staining of gels, post-separation gel manipulation, and MS
interfacing are all time-consuming tasks prone to irreproducibility, significant sample loss, and poor quantitative
accuracy. Thus, automated, high resolution, rapid, reproducible, and ultrasensitive 2D separation techniques
are needed for large-scale analysis of proteins as well as differential display of protein expression. To this end, a
microfluidic 2D protein separation platform combining isoelectric focusing and SDS gel electrophoresis will be
presented. By combining traditional 2D PAGE separations in an automated, massively-parallel microfluidic chip,
analysis cycles of minutes are now feasible, compared with hours for 2D-PAGE. The system integrates sample
handling and multidimensional separations with laser-induced fluorescence detection, enabling high sensitivity and
dynamic range with negligible analyte loss. An overview of the technology and overall system will be provided, and
applications in differential display of protein expression will be presented. Integration of an efficient chip-to-MS
interface for high throughput large-scale protein analysis will also be discussed.
9:00 am Thursday, February 5 Microchip – Separations Room A4
Stevan Jovanovich
Silicon Valley Scientific
4059 Clipper Court
Fremont, California 94538
stevan@svsci.com
77
Co-Author(s)
Iuliu Blaga, David Rank,
Norman Burns, William Gurske,
Roger McIntosh
High Throughput Preparation of Nanoscale Samples for Capillary Array Electrophoresis and a
Microchip-based Analysis System
New sample preparation and analysis technologies are required to reduce sample volumes and increase analysis
throughput for DNA sequencing, genotyping, proteomics, and drug screening. At Silicon Valley Scientific, we are
developing a microfluidics system, the NanoPrep System, to perform biochemical processing of 500 nL volumes
in arrays of capillaries in cassettes. The NanoPrep System is robust for dye-primer and dye-terminator cycle
sequencing, PCR, single-base extension, and other reactions. For DNA preparations, a template normalization
process results in readlengths and success rates that are equivalent to or better than full-volume reactions.
We describe both manual and automated versions of the NanoPrep System and their use in high throughput
sequencing using TempliPhi and advanced sample cleanup methods for capillary array electrophoresis on the
MegaBACE family of instruments and on a microchip-based analysis system.
PODIUM ABSTRACTS