omation mbers - Society for Laboratory Automation and Screening

WP066
Yu Suen
Beckman Coulter, Inc.
Biological Systems Operation
4300 N. Harbor Boulevard
Fullerton, California 92834
Ysuen@beckman.com
224
Co-Author(s)
Keith Roby
Javorka Gunic
Michael H. Simonian
Automation of Caco-2 Cell Preparation, Drug Permeation and Transport Assay on the Biomek
3000 Laboratory Automation Workstation
Incorporating predictive ADME (absorption, distribution, metabolism and elimination) assays in earlier stages
of drug discovery can help in rejecting candidate molecules that lack necessary pharmacological properties. A
human colorectal adenocarcinoma cell line, Caco-2 can be used to assess absorption, permeation and efflux
transport properties of a candidate drug. A high throughput Caco-2 assay can provide useful information for lead
optimization in the drug discovery industry. This poster describes the use of Beckman Coulter’s Biomek 3000
Laboratory Automation Workstation to automate Caco-2 cell preparation and differentiation in the BD Falcon
HTS 96-Multiwell Insert System. Additionally, Beckman Coulter’s Biomek 3000 Laboratory Automation Workstation
was used to automate the compound permeability, efflux testing and sample collection. We have demonstrated
intact and functional Caco-2 monolayers after 21 days of culture manipulation by the Biomek 3000 Laboratory
Automation Workstation. The Caco-2 monolayer provided a selective barrier for transcellular and carrier-mediated
efflux transport of different drugs. The permeability ranking of drug standards using the Caco-2 assay system
matched well with the Potential Internal Standards suggested by the FDA. The bi-directional transport studies
verified functional P-glycoprotein efflux pump activities. The Biomek 3000 Laboratory Automation Workstation
facilitated Caco-2 assay implementation, reduced the chance of contamination and minimized the intensive
requirement of sterile skills. The automated Caco-2 assay can be used for high throughput screening of drug
candidates for absorption
WP067
Joseph Suzow
Pediatrix Screening
Molecular Genetics
90 Emerson Lane
Bridgeville, Pennsylvania 15017
jsuzow@yahoo.com
Application of Automation for Clinical Newborn Screening: Detection of Hearing Loss
Associated Cytomegalovirus
Co-Author(s)
Zhili Lin
Michelle Lage
Edwin W. Naylor
This study consists of a clinical newborn screening application for the detection of cytomegalovirus (CMV). CMV
infection in the newborn is associated with the onset of hearing loss. Early detection of hearing loss is critical
for normal child development. Specimens are collected on a paper filter card. After drying, a small paper disk is
punched into each well of a 96-well plate. A Beckman Coulter Core System is then used to extract DNA from
the blood spot and set up a PCR reaction. The Core System consists of a Biomek FX liquid handler, an ORCA
arm, peripheral heat blocks, stacker carousels, plate sealer, and plate piercer. A 20µl PCR reaction is set up in
a 384 well format. The reaction consists of FRET hybridization probes combined with common PCR reagents.
Following amplification, product is detected using the Roche Light Typer instrument. Detection involves monitoring
of fluorescence during a post PCR melting cycle. A mathematical derivative of the resulting melting slope is used
to convert the slope to a peak. The apex of the peak corresponds to the melting temperature of the detection
probe. Detection of a peak indicates the presence of amplified CMV DNA. The Light Typer instrument is capable of
detecting 384 specimens in a single 10 minute run. We have demonstrated the ability to screen greater than 2000
clinical specimens each day.