omation mbers - Society for Laboratory Automation and Screening

TP019
Robin Clark
deCODE genetics
BioStructures Group
7869 NE Day Road W.
Bainbridge Island, Washington 98110
rclark@decode.com
Protein Maker: An Automated System for Protein Purification
Co-Author(s)
Alexandrina Muntianu, Hans-Thomas Richter, Denise Conner,
Lawrence Chun, Lance Stewart
The Protein Maker is an automated system for parallel liquid chromatography at medium scale (1-50 mg of
protein). Protein solutions, wash buffers and elution buffers are delivered under positive pressure to up to 24
columns by automated syringe pumps. Thus Protein Maker is analogous to an FPLC system that can run 24
columns at a time. The 24 syringe pumps are connected to 8-way valves, with each pump and valve controlled
independently through the software. A gantry with XYZ directional control carries two 24-port manifolds: one for
sample loading and one for columns. Standard pre-packed 1 to 10 ml columns are mounted on the gantry with
flow rates adjustable from 0.25 to 290 ml/min. Column fractions are delivered into 24-well block plates at any of 23
deck locations. Protein Maker can be used to purify up to 24 different proteins in parallel on duplicate columns, or
to test multiple purification strategies on one or a few proteins. Application results will be presented.
TP020
Matthew Cook
TTP LabTech
Melbourn Science Park
Melbourn, Herts SG8 6EE United Kingdom
matthew.cook@ttplabtech.com
155
Co-Author(s)
Olivier Dery, Gwyneth Olson, Annick Le Gall and
Francine Fang, TTP LabTech
Pierre Turpin, BD Bioscience Clontech
BD Biosciences Clontech ZsProSensor-1, a Fluorescent Protein-based Proteasome Sensor
Assay: Evaluation Using the Acumen Explorer
Protein degradation by the proteasome is at the core of many pathological processes such as inflammation,
autoimmunity, neurodegenerative diseases, and cancer. This has motivated efforts to identify compounds that
modulate the activity of the proteasome. Assays to monitor this activity in live cells could boost these efforts by
bypassing heavy biochemical manipulations. In this study, a proteasome targeting sequence was used to direct
the reef coral fluorescent protein ZsGreen to degradation by the proteasome. The chimaeric fluorescent protein
ZsProSensor-1 is constitutively degraded by the proteasome in stably transfected HEK 293 cells and accumulates
under conditions that alter the activity of the proteasome. Because of its high sensitivity, this live cell assay allows
monitoring of the activity of the proteasome by microscopes, flow cytometers and standard 96-well plate readers.
Using the Acumen Explorer the inhibitory activities of four different inhibitors of the proteasome were detected
at discrete time points using ZsProSensor-1-expressing cells. Propidium Iodide (PI) was used to monitor the
cytotoxicity of these compounds. These data demonstrate the complimentary nature of BD Biosciences Clontech’s
Novel Fluorescent Protein (NFP) based assays and the Acumen Explorer fluorescent detection system for the
potential identification of compounds that modulate the activity of the proteasome as well as their cytotoxicity and
other cellular effects.
POSTER ABSTRACTS